pubmed: spinal cord injury a...
NCBI: db=pubmed; Term=spinal cord injury and stem cell therapy
NCBI pubmed
  • Treatment of spinal cord injury: A review of engineering using neural and mesenchymal stem cells.

    Treatment of spinal cord injury: A review of engineering using neural and mesenchymal stem cells.

    Clin Anat. 2014 Aug 23;

    Authors: Mortazavi MM, Harmon OA, Adeeb N, Deep A, Tubbs RS

    Abstract
    Over time, various treatment modalities for spinal cord injury have been trialed, including pharmacological and nonpharmacological methods. Among these, replacement of the injured neural and paraneural tissues via cellular transplantation of neural and mesenchymal stem cells has been the most attractive. Extensive experimental studies have been done to identify the safety and effectiveness of this transplantation in animal and human models. Herein, we review the literature for studies conducted, with a focus on the human-related studies, recruitment, isolation, and transplantation, of these multipotent stem cells, and associated outcomes. Clin. Anat., 2014. © 2014 Wiley Periodicals, Inc.

    PMID: 25156268 [PubMed - as supplied by publisher]

  • SOX9 as a Predictor for Neurogenesis Potentiality of Amniotic Fluid Stem Cells.

    SOX9 as a Predictor for Neurogenesis Potentiality of Amniotic Fluid Stem Cells.

    Stem Cells Transl Med. 2014 Aug 25;

    Authors: Wei PC, Chao A, Peng HH, Chao AS, Chang YL, Chang SD, Wang HS, Chang YJ, Tsai MS, Sieber M, Chen HC, Chen SJ, Lee YS, Hwang SM, Wang TH

    Abstract
    Preclinical studies of amniotic fluid-derived cell therapy have been successful in the research of neurodegenerative diseases, peripheral nerve injury, spinal cord injury, and brain ischemia. Transplantation of human amniotic fluid stem cells (AFSCs) into rat brain ventricles has shown improvement in symptoms of Parkinson's disease and also highlighted the minimal immune rejection risk of AFSCs, even between species. Although AFSCs appeared to be a promising resource for cell-based regenerative therapy, AFSCs contain distinctly a heterogeneous pool of cell types, rendering each preparation of AFSCs unique. Identification of predictive markers for neuron-prone AFSCs is necessary before such stem cell-based therapeutics can become a reality. In an attempt to identify markers of AFSCs to predict their ability for neurogenesis, we performed a two-phase study. In the discovery phase of 23 AFSCs, we tested ZNF521/Zfp521, OCT6, SOX1, SOX2, SOX3, and SOX9 as predictive markers of AFSCs for neural differentiation. In the validation phase, the efficacy of these predictive markers was tested in independent sets of 18 AFSCs and 14 dental pulp stem cells (DPSCs). We found that high expression of SOX9 in AFSCs is associated with good neurogenetic ability, and these positive correlations were confirmed in independent sets of AFSCs and DPSCs. Furthermore, knockdown of SOX9 in AFSCs inhibited their neuronal differentiation. In conclusion, the discovery of SOX9 as a predictive marker for neuron-prone AFSCs could expedite the selection of useful clones for regenerative medicine, in particular, in neurological diseases and injuries.

    PMID: 25154783 [PubMed - as supplied by publisher]

  • Fibroblasts isolated from human middle turbinate mucosa cause neural progenitor cells to differentiate into glial lineage cells.
    Related Articles

    Fibroblasts isolated from human middle turbinate mucosa cause neural progenitor cells to differentiate into glial lineage cells.

    PLoS One. 2013;8(10):e76926

    Authors: Wu X, Bolger WE, Anders JJ

    Abstract
    Transplantation of olfactory ensheathing cells (OECs) is a potential therapy for repair of spinal cord injury (SCI). Autologous transplantation of OECs has been reported in clinical trials. However, it is still controversial whether purified OECs or olfactory mucosa containing OECs, fibroblasts and other cells should be used for transplantation. OECs and fibroblasts were isolated from olfactory mucosa of the middle turbinate from seven patients. The percentage of OECs with p75(NTR+) and GFAP(+) ranged from 9.2% to 73.2%. Fibroblasts were purified and co-cultured with normal human neural progenitors (NHNPs). Based on immunocytochemical labeling, NHNPs were induced into glial lineage cells when they were co-cultured with the mucosal fibroblasts. These results demonstrate that OECs can be isolated from the mucosa of the middle turbinate bone as well as from the dorsal nasal septum and superior turbinates, which are the typical sites for harvesting OECs. Transplantation of olfactory mucosa containing fibroblasts into the central nervous system (CNS) needs to be further investigated before translation to clinical application.

    PMID: 24204706 [PubMed - indexed for MEDLINE]

pubmed: sports injury and st...
NCBI: db=pubmed; Term=sports injury and stem cell therapy
NCBI pubmed
  • Mesenchymal stem cells in synovial fluid increase after meniscus injury.
    Related Articles

    Mesenchymal stem cells in synovial fluid increase after meniscus injury.

    Clin Orthop Relat Res. 2014 May;472(5):1357-64

    Authors: Matsukura Y, Muneta T, Tsuji K, Koga H, Sekiya I

    Abstract
    BACKGROUND: Although relatively uncommon, spontaneous healing from a meniscus injury has been observed even within the avascular area. This may be the result of the existence of mesenchymal stem cells in synovial fluid.
    QUESTIONS/PURPOSES: The purpose of this study was to investigate whether mesenchymal stem cells existed in the synovial fluid of the knee after meniscus injury.
    METHODS: Synovial fluid was obtained from the knees of 22 patients with meniscus injury just before meniscus surgery and from 8 volunteers who had no history of knee injury. The cellular fraction of the synovial fluid was cultured for 14 days followed by analysis for multilineage potential and presentation of surface antigens characteristic of mesenchymal stem cells. Colony-forming efficiency and proliferation potential were also compared between the two groups.
    RESULTS: Cells with characteristics of mesenchymal stem cells were observed in the synovial fluid of injured knees to a much greater degree than in uninjured knees. The colony-forming cells derived from the synovial fluid of the knee with meniscus injury had multipotentiality and surface epitopes identical to mesenchymal stem cells. The average number of colony formation, obtained from 1 mL of synovial fluid, in meniscus-injured knees was 250, higher than that from healthy volunteers, which was 0.5 (p < 0.001). Total colony number per synovial fluid volume was positively correlated with the postinjury period (r = 0.77, p < 0.001).
    CONCLUSIONS: Mesenchymal stem cells were found to exist in synovial fluid from knees after meniscus injury. Mesenchymal stem cells were present in higher numbers in synovial fluid with meniscus injury than in normal knees. Total colony number per synovial fluid volume was positively correlated with the postinjury period.
    CLINICAL RELEVANCE: Our current human study and previous animal studies suggest the possibility that mesenchymal stem cells in synovial fluid increase after meniscus injury contributing to spontaneous meniscus healing.

    PMID: 24338094 [PubMed - indexed for MEDLINE]

Thrombospondin-1 is a plasmatic marker of peripheral arterial disease that modulates endothelial progenitor cell angiogenic properties.
Source:  NIH.gov
Monday, 29 November 1999 17:00

Thrombospondin-1 is a plasmatic marker of peripheral arterial disease that modulates endothelial progenitor cell angiogenic properties.

Arterioscler Thromb Vasc Biol. 2011 Mar;31(3):551-9

Authors: Smadja DM, d'Audigier C, Bièche I, Evrard S, Mauge L, Dias JV, Labreuche J, Laurendeau I, Marsac B, Dizier B, Wagner-Ballon O, Boisson-Vidal C, Morandi V, Duong-Van-Huyen JP, Bruneval P, Dignat-George F, Emmerich J, Gaussem P

Abstract
OBJECTIVE: We examined whether plasma levels of angiogenic factors are altered in plasma of patients with peripheral arterial disease (PAD) and whether these factors affect endothelial progenitor cell-induced angiogenesis.
METHODS AND RESULTS: Plasma was collected from 184 patients with PAD and 330 age-matched healthy controls. Vascular endothelial growth factor and placental growth factor concentrations did not differ between the groups, whereas we found a linear correlation between PAD disease and thrombospondin (TSP)-1 plasma level. TSP-1 was expressed in newly formed vessels in PAD patients having received local injections of bone marrow mononuclear cells. To analyze the functional role of TSP-1 during neoangiogenesis, we used a Matrigel-plug assay and showed that vascularization of implanted Matrigel-plugs was increased in TSP-1(-/-) mice. Moreover, injections of TSP-1 in C57Bl6/J mice after hindlimb ischemia induced a significant decrease of blood flow recovery. To investigate the effects of TSP-1 on human endothelial colony-forming cell (ECFC) angiogenic potential, recombinant human TSP-1 and a small interfering RNA were used. In vitro, TSP-1 N-terminal part significantly enhanced ECFC adhesion, whereas recombinant human TSP-1 had a negative effect on ECFC angiogenic potential. This effect, mediated by CD47 binding, modulated stromal cell-derived factor 1/CXC chemokine receptor 4 pathway.
CONCLUSIONS: TSP-1 is a potential biomarker of PAD and ECFC-induced angiogenesis, suggesting that TSP-1 modulation might improve local tissue ischemia in this setting. (Clinical trial registration: NCT00377897.).

PMID: 21148423 [PubMed - indexed for MEDLINE]


Synergistic effect of adipose-derived stem cell therapy and bone marrow progenitor recruitment in ischemic heart.
Source:  NIH.gov
Monday, 29 November 1999 17:00

Synergistic effect of adipose-derived stem cell therapy and bone marrow progenitor recruitment in ischemic heart.

Lab Invest. 2011 Apr;91(4):539-52

Authors: Ii M, Horii M, Yokoyama A, Shoji T, Mifune Y, Kawamoto A, Asahi M, Asahara T

Abstract
Human multipotent adipose-derived stem cells (hMADSCs) have recently been isolated featuring extensive expansion capacity ex vivo. We tested the hypothesis that hMADSC transplantation might contribute to cardiac functional recovery by its direct or indirect effect on myocardial infarction (MI). Nude rats were either transplanted with hMADSCs or PBS (control) in ischemic myocardium immediately following MI. Echocardiographical assessment of cardiac function after MI with hMADSCs showed significant improvement of each parameter compared to that with PBS. Histological analysis also showed significantly reduced infarct size and increased capillary density in peri-infarct myocardium by hMADSC treatment. However, remarkable transdifferentiation of hMADSCs into cardiac or vascular lineage cells was not observed. Despite the less transdifferentiation capacity, hMADSCs produced robust multiple pro-angiogenic growth factors and chemokines, such as vascular endothelial growth factor (VEGF), basic fibroblast growth factor (bFGF), and stromal cell-derived factor-1α (SDF-1α). Specifically, hMADSC-derived SDF-1α had a crucial role for cooperative angiogenesis, with the paracrine effect of hMADSCs and Tie2-positive bone marrow (BM) progenitor recruitment in ischemic myocardium. hMADSCs exhibit a therapeutic effect on cardiac preservation following MI, with the production of VEGF, bFGF, and SDF-1α showing paracrine effects and endogenous BM stem/progenitor recruitment to ischemic myocardium rather than its direct contribution to tissue regeneration.

PMID: 21135814 [PubMed - indexed for MEDLINE]


Pro-angiogenic induction of myeloid cells for therapeutic angiogenesis can induce mitogen-activated protein kinase p38-dependent foam cell formation.
Source:  NIH.gov
Monday, 29 November 1999 17:00

Pro-angiogenic induction of myeloid cells for therapeutic angiogenesis can induce mitogen-activated protein kinase p38-dependent foam cell formation.

Cytotherapy. 2011 Apr;13(4):503-12

Authors: Rohde E, Schallmoser K, Reinisch A, Hofmann NA, Pfeifer T, Fröhlich E, Rechberger G, Lanzer G, Kratky D, Strunk D

Abstract
BACKGROUND AIMS: Clinical trials for therapeutic angiogenesis use blood- or bone marrow-derived hematopoietic cells, endothelial progenitor cells (EPC) and mesenchymal stromal cells (MSC) for vascular regeneration. Recently concerns have emerged that all three cell types could also contribute to atherosclerosis by foam cell formation. Therefore, we asked whether human myelomonocytic cells, EPC or MSC can accumulate lipid droplets (LD) and develop into foam cells.
METHODS: LD accumulation was quantified by flow cytometry, confocal microscopy and cholesterol measurement in each of the cell types. The impact of an initial pro-angiogenic induction on subsequent foam cell formation was studied to mimic relevant settings already used in clinical trials. The phosphorylation state of intracellular signaling molecules in response to the pro-angiogenic stimulation was determined to delineate the operative mechanisms and establish a basis for interventional strategies.
RESULTS: Foam cells were formed by monocytes but not by EPC or MSC after pro-angiogenic induction. Mitogen-activated protein kinase (MAPK) p38 phosphorylation was enhanced and kinase inhibition almost abrogated intracellular LD accumulation in monocytes.
CONCLUSIONS: These data suggest that hematopoietic cell preparations containing monocytes bear the risk of foam cell formation after pro-angiogenic induction. Instead, EPC and MSC may drive vascular regeneration without atherogenesis aggravation. A thorough understanding of cell biology is necessary to develop new strategies combining pro-angiogenic and anti-atherogenic effects during cell therapy.

PMID: 21128706 [PubMed - indexed for MEDLINE]


Nonexpanded mesenchymal stem cells for regenerative medicine: yield in stromal vascular fraction from adipose tissues.
Source:  NIH.gov
Monday, 29 November 1999 17:00

Nonexpanded mesenchymal stem cells for regenerative medicine: yield in stromal vascular fraction from adipose tissues.

Tissue Eng Part C Methods. 2010 Dec;16(6):1515-21

Authors: Faustini M, Bucco M, Chlapanidas T, Lucconi G, Marazzi M, Tosca MC, Gaetani P, Klinger M, Villani S, Ferretti VV, Vigo D, Torre ML

Abstract
The adipose-derived stromal vascular fraction (SVF) represents a rich source of mesenchymal cells, potentially able to differentiate into adipocytes, chondrocytes, osteoblasts, myocytes, cardiomyocytes, hepatocytes, and neuronal, epithelial, and endothelial cells. These cells are ideal candidates for use in regenerative medicine, tissue engineering, including gene therapy, and cell replacement cancer therapies. In this work, we aimed to the optimization of the adipose SVF-based therapy, and the effect of the collection site, surgical procedure, and tissue processing techniques on SVF yield was evaluated in terms of cell recovery and live cells, taking into account the effect of gender, age, and body mass index. Adipose tissue samples were recovered from 125 informed subjects (37 males and 88 females; mean age: 51.31 years; range: 15-87 years), and digested in different condition with collagenase. A multivariate linear model put in evidence that in males the best collection site in terms of yield is located in the abdomen, whereas in females the biopsy region do not influence cell recovery; the collection technique, the age, and the body mass index of donor seem not to influence the cell yield. The tissue-processing procedures strongly modify the yield and the vitality of cells: a collagenase concentration of 0.2% and a digestion time of 1 h could be chosen as the best operating conditions.

PMID: 20486782 [PubMed - indexed for MEDLINE]


PI3K/p110{delta} is a novel therapeutic target in multiple myeloma.
Source:  NIH.gov
Monday, 29 November 1999 17:00

PI3K/p110{delta} is a novel therapeutic target in multiple myeloma.

Blood. 2010 Sep 2;116(9):1460-8

Authors: Ikeda H, Hideshima T, Fulciniti M, Perrone G, Miura N, Yasui H, Okawa Y, Kiziltepe T, Santo L, Vallet S, Cristea D, Calabrese E, Gorgun G, Raje NS, Richardson P, Munshi NC, Lannutti BJ, Puri KD, Giese NA, Anderson KC

Abstract
In this study, we demonstrate expression and examined the biologic sequelae of PI3K/p110delta signaling in multiple myeloma (MM). Knockdown of p110delta by small interfering RNA caused significant inhibition of MM cell growth. Similarly, p110delta specific small molecule inhibitor CAL-101 triggered cytotoxicity against LB and INA-6 MM cell lines and patient MM cells, associated with inhibition of Akt phosphorylation. In contrast, CAL-101 did not inhibit survival of normal peripheral blood mononuclear cells. CAL-101 overcame MM cell growth conferred by interleukin-6, insulin-like growth factor-1, and bone marrow stromal cell coculture. Interestingly, inhibition of p110delta potently induced autophagy. The in vivo inhibition of p110delta with IC488743 was evaluated in 2 murine xenograft models of human MM: SCID mice bearing human MM cells subcutaneously and the SCID-hu model, in which human MM cells are injected within a human bone chip implanted subcutaneously in SCID mice. IC488743 significantly inhibited tumor growth and prolonged host survival in both models. Finally, combined CAL-101 with bortezomib induced synergistic cytotoxicity against MM cells. Our studies therefore show that PI3K/p110delta is a novel therapeutic target in MM and provide the basis for clinical evaluation of CAL-101 to improve patient outcome in MM.

PMID: 20505158 [PubMed - indexed for MEDLINE]


Unsorted human adipose tissue-derived stem cells promote angiogenesis and myogenesis in murine ischemic hindlimb model.
Source:  NIH.gov
Monday, 29 November 1999 17:00

Unsorted human adipose tissue-derived stem cells promote angiogenesis and myogenesis in murine ischemic hindlimb model.

Microvasc Res. 2010 Dec;80(3):310-6

Authors: Kang Y, Park C, Kim D, Seong CM, Kwon K, Choi C

Abstract
We examined the protective effect of unsorted human adipose tissue-derived stem cells (hADSCs) with a short-term culture in endothelial differentiation medium on tissue repair after ischemic injury. hADSCs were isolated from human subcutaneous adipose tissue and cultured in vitro in endothelial differentiation medium for 2wks before transplantation. Cultured hADSCs showed a typical mesenchymal stromal cell-like phenotype, positive for endothelial-specific markers including VE-cadherin, Flt-1, eNOS, and vWF but not CD31. Two hours after ligation of the femoral artery and vein, mice were injected with the unselected hADSCs locally near the surgery site and tested for tissue perfusion and repair. Tissue perfusion rates of the ischemic limbs were significantly higher in the group treated with hADSCs compared with those of the control mice as early as post-operative day 3 (median 195.3%/min; interquartile range, 82.0-321.1 vs. median 47.1%/min; interquartile range, 18.0-58.7; p=0.001 by Friedman two-way analysis). Subsequently, the mice treated with hADSC showed better prognosis at 4wks after surgery, and the histological analysis revealed increased vascular density and reduced muscle atrophy in the hADSC-transplanted limbs. Moreover, hADSC-treated muscle contained differentiated myocytes positive for human NF-κB and myogenin antigen. These results collectively indicate that unsorted hADSCs after a 2-wk-in vitro culture have a therapeutic potential in ischemic tissue injury via inducing both angiogenesis and myogenesis.

PMID: 20510252 [PubMed - indexed for MEDLINE]


A long-term follow-up study of intravenous autologous mesenchymal stem cell transplantation in patients with ischemic stroke.
Source:  NIH.gov
Monday, 29 November 1999 17:00

A long-term follow-up study of intravenous autologous mesenchymal stem cell transplantation in patients with ischemic stroke.

Stem Cells. 2010 Jun;28(6):1099-106

Authors: Lee JS, Hong JM, Moon GJ, Lee PH, Ahn YH, Bang OY,

Abstract
We previously evaluated the short-term follow-up preliminary data of mesenchymal stem cells (MSCs) transplantation in patients with ischemic stroke. The present study was conducted to evaluate the long-term safety and efficacy of i.v. MSCs transplantation in a larger population. To accomplish this, we performed an open-label, observer-blinded clinical trial of 85 patients with severe middle cerebral artery territory infarct. Patients were randomly allocated to one of two groups, those who received i.v. autologous ex vivo cultured MSCs (MSC group) or those who did not (control group), and followed for up to 5 years. Mortality of any cause, long-term side effects, and new-onset comorbidities were monitored. Of the 52 patients who were finally included in this study, 16 were the MSC group and 36 were the control group. Four (25%) patients in the MSC group and 21 (58.3%) in the control group died during the follow-up period, and the cumulative surviving portion at 260 weeks was 0.72 in the MSC group and 0.34 in the control group (log-rank; p = .058). Significant side effects were not observed following MSC treatment. The occurrence of comorbidities including seizures and recurrent vascular episodes did not differ between groups. When compared with the control group, the follow-up modified Rankin Scale (mRS) score was decreased, whereas the number of patients with a mRS of 0-3 increased in the MSC group (p = .046). Clinical improvement in the MSC group was associated with serum levels of stromal cell-derived factor-1 and the degree of involvement of the subventricular region of the lateral ventricle. Intravenous autologous MSCs transplantation was safe for stroke patients during long-term follow-up. This therapy may improve recovery after stroke depending on the specific characteristics of the patients.

PMID: 20506226 [PubMed - indexed for MEDLINE]


In vivo evaluation of mixtures of uncultured freshly isolated adipose-derived stem cells and demineralized bone matrix for bone regeneration in a rat critically sized calvarial defect model.
Source:  NIH.gov
Monday, 29 November 1999 17:00

In vivo evaluation of mixtures of uncultured freshly isolated adipose-derived stem cells and demineralized bone matrix for bone regeneration in a rat critically sized calvarial defect model.

Stem Cells Dev. 2011 Feb;20(2):233-42

Authors: Rhee SC, Ji YH, Gharibjanian NA, Dhong ES, Park SH, Yoon ES

Abstract
Although many studies have suggested that human adipose tissue contains pluripotent stem cells, a few reports are available on stromal vascular fraction (SVF). In the present study, we evaluated the bone formation capacities of SVF. We implanted uncultured freshly isolated adipose-derived stem cells combined with demineralized bone matrix (DBM) to induce bone regeneration in a critically sized rat calvarial defect model. We used DBM (DBX(®)) and/or poly(70L-lactide-co-30DL-lactide) copolymer PLA as a scaffold. Fifty white rats were randomized to 5 different groups (n=10): (1) control, (2) DBM, (3) DBM + SVF, (4) DBM + PLA, and (5) DBM + PLA + SVF groups. After acquiring SVF, an 8-mm critically sized calvarial defect was made in each rat. Specimens were harvested at 8 weeks postimplantation and evaluated radiographically and histologically. New bone formation was qualified by hematoxylin and eosin staining and anti-osteocalcin antibody (OC4-30) immunostaining of calvarial sections. Amounts of mineralization were determined by radiodensitometric analysis. In gross appearance, the DBM + SVF and DBM + PLA + SVF groups showed more abundant bone formation than the other groups. Radiodensitometric evaluations revealed that significant intergroup differences were observed according to the Kruskal-Wallis (rank) test (P=0.030<0.05). The 5 groups show different amounts of filling of bone defects (control: 13.48%; DBM: 39.94%; DBM + SVF: 57.69%; DBM + PLA: 24.86%; DBM + PLA + SVF: 42.75%). Histological evaluation revealed that there was abundant new bone formation in the DBM + SVF and DBM + PLA + SVF groups. It was found that undifferentiated adipose-derived stem cells in the form of SVF induced new bone formation in rat calvarial defects. Accordingly, SVF offers a practical, promising candidate for regenerative tissue engineering or cell-based therapy.

PMID: 20528145 [PubMed - indexed for MEDLINE]


Autologous stromal vascular fraction cells: a tool for facilitating tolerance in rheumatic disease.
Source:  NIH.gov
Monday, 29 November 1999 17:00

Autologous stromal vascular fraction cells: a tool for facilitating tolerance in rheumatic disease.

Cell Immunol. 2010;264(1):7-17

Authors: Ichim TE, Harman RJ, Min WP, Minev B, Solano F, Rodriguez JP, Alexandrescu DT, De Necochea-Campion R, Hu X, Marleau AM, Riordan NH

Abstract
Since the days of Medawar, the goal of therapeutic tolerogenesis has been a "Holy Grail" for immunologists. While knowledge of cellular and molecular mechanisms of this process has been increasing at an exponential rate, clinical progress has been minimal. To provide a mechanistic background of tolerogenesis, we overview common processes in the naturally occurring examples of: pregnancy, cancer, oral tolerance and anterior chamber associated immune deviation. The case is made that an easily accessible byproduct of plastic surgery, the adipose stromal vascular fraction, contains elements directly capable of promoting tolerogenesis such as T regulatory cells and inhibitory macrophages. The high content of mesenchymal and hematopoietic stem cells from this source provides the possibility of trophic/regenerative potential, which would augment tolerogenic processes by decreasing ongoing inflammation. We discuss the application of this autologous cell source in the context of rheumatoid arthritis, concluding with some practical examples of its applications.

PMID: 20537320 [PubMed - indexed for MEDLINE]


Glycolytic cancer associated fibroblasts promote breast cancer tumor growth, without a measurable increase in angiogenesis: evidence for stromal-epithelial metabolic coupling.
Source:  NIH.gov
Monday, 29 November 1999 17:00

Glycolytic cancer associated fibroblasts promote breast cancer tumor growth, without a measurable increase in angiogenesis: evidence for stromal-epithelial metabolic coupling.

Cell Cycle. 2010 Jun 15;9(12):2412-22

Authors: Migneco G, Whitaker-Menezes D, Chiavarina B, Castello-Cros R, Pavlides S, Pestell RG, Fatatis A, Flomenberg N, Tsirigos A, Howell A, Martinez-Outschoorn UE, Sotgia F, Lisanti MP

Abstract
Previously, we proposed a new model for understanding the Warburg effect in tumorigenesis and metastasis. In this model, the stromal fibroblasts would undergo aerobic glycolysis (a.k.a., the Warburg effect)--producing and secreting increased pyruvate/lactate that could then be used by adjacent epithelial cancer cells as "fuel" for the mitochondrial TCA cycle, oxidative phosphorylation, and ATP production. To test this model more directly, here we used a matched set of metabolically well-characterized immortalized fibroblasts that differ in a single gene. CL3 fibroblasts show a shift towards oxidative metabolism, and have an increased mitochondrial mass. In contrast, CL4 fibroblasts show a shift towards aerobic glycolysis, and have a reduced mitochondrial mass. We validated these differences in CL3 and CL4 fibroblasts by performing an unbiased proteomics analysis, showing the functional upregulation of 4 glycolytic enzymes, namely ENO1, ALDOA, LDHA and TPI1, in CL4 fibroblasts. Many of the proteins that were upregulated in CL4 fibroblasts, as seen by unbiased proteomics, were also transcriptionally upregulated in the stroma of human breast cancers, especially in the patients that were prone to metastasis. Importantly, when CL4 fibroblasts were co-injected with human breast cancer cells (MDA-MB-231) in a xenograft model, tumor growth was dramatically enhanced. CL4 fibroblasts induced a > 4-fold increase in tumor mass, and a near 8-fold increase in tumor volume, without any measurable increases in tumor angiogenesis. In parallel, CL3 and CL4 fibroblasts both failed to form tumors when they were injected alone, without epithelial cancer cells. Mechanistically, under co-culture conditions, CL4 glycolytic fibroblasts increased mitochondrial activity in adjacent breast cancer cells (relative to CL3 cells), consistent with the "Reverse Warburg Effect". Notably, Western blot analysis of CL4 fibroblasts revealed a significant reduction in caveolin-1 (Cav-1) protein levels. In human breast cancer patients, a loss of stromal Cav-1 is associated with an increased risk of early tumor recurrence, metastasis, tamoxifen-resistance, and poor clinical outcome. Thus, loss of stromal Cav-1 may be an effective marker for predicting the "Reverse Warburg Effect" in the stroma of human breast cancer patients. As such, CL4 fibroblasts are a new attractive model for mimicking the "glycolytic phenotype" of cancer-associated fibroblasts. Nutrients derived from glycolytic cancer associated fibroblasts could provide an escape mechanism to confer drug-resistance during anti-angiogenic therapy, by effectively reducing the dependence of cancer cells on a vascular blood supply.

PMID: 20562527 [PubMed - in process]


Integrin expression and integrin-mediated adhesion in vitro of human multipotent stromal cells (MSCs) to endothelial cells from various blood vessels.
Source:  NIH.gov
Monday, 29 November 1999 17:00

Integrin expression and integrin-mediated adhesion in vitro of human multipotent stromal cells (MSCs) to endothelial cells from various blood vessels.

Cell Tissue Res. 2010 Jul;341(1):147-58

Authors: Semon JA, Nagy LH, Llamas CB, Tucker HA, Lee RH, Prockop DJ

Abstract
Multipotent mesenchymal stromal cells (MSCs) home to damaged tissue by processes partly regulated by integrins. Integrin subunits expressed by MSCs were identified by flow cytometry (FC), immunocytochemistry (IC), and a panel of integrin-binding antibodies. In subconfluent cultures, over 80% of MSCs expressed integrin subunits beta1, beta2, and alpha3, 20%-55% expressed alpha1, alpha2, alpha4, alpha5, alpha6, and alphaV, and about 10% expressed beta3 when assayed by FC. None of the cells expressed significant levels of 13 other integrins as assayed by FC, but seven of the 13 integrins were detected by IC: beta5, alpha7, alpha8, alpha9, alpha11, alphaX, and alphaD. Expression of some integrins changed with MSC confluency: integrins beta3, alpha1, alpha3, alpha5, and alphaV increased, and alpha6 decreased. Furthermore, alpha4 was the only integrin to vary among preparations of MSCs from different donors. The results resolved some discrepancies in the literature concerning integrin expression by MSCs. We also investigated the role of specific integrins in MSC adhesion to endothelial cells (ECs) from the pulmonary artery (HPAEC), cardiac-derived microvasculature (HMVEC-C), and umbilical veins (HUVEC). In experiments with blocking antibodies to beta integrins, anti-beta5 reduced MSC adhesion to all types of ECs, anti-beta1 to both HUVEC and HPAEC, anti-beta3 to HUVEC, and anti-beta2 to HMVEC-C. With blocking antibodies to alpha integrins, anti-alphaX reduced adhesion to HPAEC and HMVEC-C, anti-alphaV to HPAEC, and both anti-alpha7 and anti-alphaD to HMVEC-C. Thus, MSCs use diverse integrins to adhere to EC from various blood vessels in vitro.

PMID: 20563599 [PubMed - indexed for MEDLINE]


Bone marrow mesenchymal stem cells: historical overview and concepts.
Source:  NIH.gov
Monday, 29 November 1999 17:00

Bone marrow mesenchymal stem cells: historical overview and concepts.

Hum Gene Ther. 2010 Sep;21(9):1045-56

Authors: Charbord P

Abstract
This review describes the historical emergence of the concept of bone marrow mesenchymal stem cells (MSCs), summarizing data on Wolf and Trentin's hematopoietic inductive microenvironment; Dexter's hematopoiesis-supportive stromal cells; Friedenstein's osteogenic cells; and Pittenger's trilineal osteoblastic, chondrocytic, and adipocytic precursors; to finally introduce the specific bone marrow mesenchymal stem cells with differentiation potential to four lineages (mesenchymal and vascular smooth muscle lineages), and stromal and immunomodulatory capacities. Two points are the object of detailed discussion. The first point envisions the stem cell attributes (multipotentiality, self-renewal, tissue regeneration, population heterogeneity, plasticity, and lineage priming) compared with that of the paradigmatic hematopoietic stem cell. In the second point, we discuss the possible existence of bone marrow cells with greater differentiation potential, eventually pluripotential cells. The latter point raises the issues of cell fusion, reprogramming, or selection under nonstandardized conditions of rare populations of neuroectodermal origin, or of cells that had undergone mesenchymal-to-epithelial transition. In the last section, we review data on MSC senescence and possible malignant transformation secondary to extensive culture, gene transfer of telomerase, or mutations such as leading to Ewing's sarcoma. The set of data leads to the conclusion that bone marrow MSCs constitute a specific adult tissue stem cell population. The multiple characteristics of this stem cell type account for the versatility of the mechanisms of injured tissue repair. Although MSC administration may be extremely useful in a number of clinical applications, their transplantation is not without risks that must not be overlooked when developing cell therapy protocols.

PMID: 20565251 [PubMed - indexed for MEDLINE]


Beneficial effects of VEGF secreted from stromal cells in supporting endothelial cell functions: therapeutic implications for critical limb ischemia.
Source:  NIH.gov
Monday, 29 November 1999 17:00

Beneficial effects of VEGF secreted from stromal cells in supporting endothelial cell functions: therapeutic implications for critical limb ischemia.

Cell Transplant. 2010;19(11):1425-37

Authors: Cobellis G, Maione C, Botti C, Coppola A, Silvestroni A, Lillo S, Schiavone V, Molinari AM, Sica V

Abstract
Critical limb ischemia (CLI) is the end stage of peripheral vascular disease (PVD). One third of CLI patients progresses to leg amputation with high associated morbidity and mortality. In no-option patients with end-stage critical limb ischemia, bone marrow cell transplantation has shown promising results, improving leg perfusion to the level of reducing major amputations and allowing limb salvage. We recently reported the successful application of an innovative protocol based on repeated autologous bone marrow cell transplantation, which resulted in an effective and feasible strategy for achieving long-term revascularization in patients with severe CLI. In an effort to understand the clinical benefit provided by stem cells therapy in patients with CLI, we characterized the marrow-derived stromal cells of CLI patients and we provided a correlation between the in vitro features of these cells and the clinical follow up at 12 months. We showed that cells derived from CLI patients had a reduced capacity to proliferate, adhere, and migrate, but that they stimulated proliferation and migration of endothelial cells through the release of VEGF-A, supporting the idea that the paracrine mechanisms underpinned the biological effects of long-term angiogenesis in CLI patients.

PMID: 20587143 [PubMed - indexed for MEDLINE]


Novel markers of mesenchymal stem cells defined by genome-wide gene expression analysis of stromal cells from different sources.
Source:  NIH.gov
Monday, 29 November 1999 17:00

Novel markers of mesenchymal stem cells defined by genome-wide gene expression analysis of stromal cells from different sources.

Exp Cell Res. 2010 Oct 1;316(16):2609-17

Authors: Kaltz N, Ringe J, Holzwarth C, Charbord P, Niemeyer M, Jacobs VR, Peschel C, Häupl T, Oostendorp RA

Abstract
Mesenchymal stem cells (MSC) represent a mixture of different cell types, of which only a minority is therapeutically relevant. Surface markers specifically identifying non-differentiated MSC from their differentiated progeny have not been described in sufficient detail. We here compare the gene expression profile of the in vivo bone-forming bone marrow-derived MSC (BM-MSC) with non-bone-forming umbilical vein stromal cells (UVSC) and other non-MSC. Clustering analysis shows that UVSC are a lineage homogeneous cell population, clearly distinct from MSC, other mesenchymal lineages and hematopoietic cells. We find that 89 transcripts of membrane-associated proteins are represented more in cultured BM-MSC than in UVSC. These include previously identified molecules, but also novel markers like NOTCH3, JAG1, and ITGA11. We show that the latter three molecules are also expressed on fibroblast colony-forming units (CFU-F). Both NOTCH3 and ITGA11, but not JAG1, further enrich for CFU-F when combined with CD146, a known marker of cells with MSC activity in vivo. Differentiation studies show that NOTCH3+ and CD146+ NOTCH3+ cells sorted from cultured BM-MSC are capable of adipogenic and osteogenic progeny, while ITGA11-expressing cells mainly show an osteogenic differentiation profile with limited adipogenic differentiation. Our observations may facilitate the study of lineage relationships in MSC as well as facilitate the development of more homogeneous cell populations for mesenchymal cell therapy.

PMID: 20599957 [PubMed - indexed for MEDLINE]


Cancer: Chemotherapy counteracted.
Source:  NIH.gov
Monday, 29 November 1999 17:00

Cancer: Chemotherapy counteracted.

Nature. 2010 Dec 2;468(7324):637-8

Authors: Emmenegger U, Kerbel RS

PMID: 21124441 [PubMed - indexed for MEDLINE]


Urokinase receptor mediates mobilization, migration, and differentiation of mesenchymal stem cells.
Source:  NIH.gov
Monday, 29 November 1999 17:00

Urokinase receptor mediates mobilization, migration, and differentiation of mesenchymal stem cells.

Cardiovasc Res. 2011 Apr 1;90(1):113-21

Authors: Vallabhaneni KC, Tkachuk S, Kiyan Y, Shushakova N, Haller H, Dumler I, Eden G

Abstract
AIMS: Multipotent mesenchymal stem cells (MSCs) have regenerative properties and are recognized as putative players in the pathogenesis of cardiovascular diseases. The underlying molecular mechanisms remain, however, sparsely explored. Our study was designed to elucidate a probable role for the multifunctional urokinase (uPA)/urokinase receptor (uPAR) system in MSC regulation. Though uPAR has been implicated in a broad spectrum of pathophysiological processes, nothing is known about uPAR in MSCs.
METHODS AND RESULTS: uPAR was required to mobilize MSCs from the bone marrow (BM) of mice stimulated with granulocyte colony-stimulating factor (G-CSF) in vivo. An insignificant amount of MSCs was mobilized in uPAR(-/-) C57BL/6J mice, whereas in wild-type animals G-CSF induced an eight-fold increase of mobilized MSCs. uPAR(-/-) mice revealed up-regulated expression of G-CSF and stromal cell-derived factor 1 (CXCR4) receptors in BM. uPAR down-regulation leads to inhibition of human MSC migration, as shown in different migration assays. uPAR down- or up-regulation resulted in inhibition or stimulation of MSC differentiation into vascular smooth muscle cells (VSMCs) correspondingly, as monitored by changes in cell morphology and expression of specific marker proteins. Injection of fluorescently labelled MSCs in non-obese diabetic/severe combined immunodeficiency (NOD/SCID) mice after femoral artery wire injury demonstrated impaired engraftment of uPAR-deficient MSCs at the place of injury.
CONCLUSIONS: These data suggest a multifaceted function of uPAR in MSC biology contributing to vascular repair. uPAR might guide and control the trafficking of MSCs to the vascular wall in response to injury or ischaemia and their differentiation towards functional VSMCs at the site of arterial injury.

PMID: 21088115 [PubMed - indexed for MEDLINE]


Short- and long-term effects of erythropoietin treatment on endothelial progenitor cell levels in patients with cardiorenal syndrome.
Source:  NIH.gov
Monday, 29 November 1999 17:00

Short- and long-term effects of erythropoietin treatment on endothelial progenitor cell levels in patients with cardiorenal syndrome.

Heart. 2011 Jan;97(1):60-5

Authors: Jie KE, van der Putten K, Bergevoet MW, Doevendans PA, Gaillard CA, Braam B, Verhaar MC

Abstract
OBJECTIVE: Patients with cardiorenal syndrome (CRS) have high cardiovascular morbidity. Endothelial progenitor cells (EPC) constitute an endogenous vascular repairsystem, protecting against atherosclerosis development. Erythropoietin (EPO) treatment may have beneficial effects by mobilizing EPC from the bonemarrow. Our objective is to determine EPC levels and effects of EPO therapy on EPC levels in CRS patients.
DESIGN: Open-label randomized trial.
SETTING: Part of the EPOCARES-trial, conducted in Utrecht (Netherlands).
PATIENTS: Patients with CRS and anaemia and healthy controls were included. Interventions Patients were randomized to receive EPO therapy (50 IU/kg/wk) for 52 weeks or no EPO therapy.
MAIN OUTCOME MEASURES: CD34(+)KDR(+)-EPC, cultured EPC outgrowth and function at baseline, after 18 days and after 52 weeks.
RESULTS: Patients showed lower CD34(+)KDR(+)-cell numbers compared to controls (6(12) vs. 19(19) cells/10(5) granulocytes; p = 0.010), despite increased levels of stromal cell-derived factor-1α; (3.1(0.8) vs 2.6(0.3) ng/ml; p = 0.001). EPC outgrowth and function were not different between patients and controls. EPC levels did not change after 18&emsp14;days with or without EPO treatment. CD34(+)KDR(+)-cells significantly declined after 52 weeks in the non-treated group (p = 0.028). Long-term EPO therapy did not significantly affect this reduction in CD34(+)KDR(+)-EPC levels.
CONCLUSIONS: CRS patients showed reduced CD34(+)KDR(+)-EPC levels compared to controls, consistent with a reduced vascular regenerative potential and despite upregulated SDF-1α levels. Over a one-year follow-up period a marked 68% further reduction in EPC levels was observed in the patient group without EPO treatment. In spite of promising experimental studies, our longitudinal, randomized study did not show significant influence of either short- or long-term EPO therapy on reduced EPC levels in CRS patients.

PMID: 21071558 [PubMed - indexed for MEDLINE]


Mesenchymal stem cells for cardiac cell therapy.
Source:  NIH.gov
Monday, 29 November 1999 17:00

Mesenchymal stem cells for cardiac cell therapy.

Hum Gene Ther. 2011 Jan;22(1):3-17

Authors: Choi YH, Kurtz A, Stamm C

Abstract
Despite refinements of medical and surgical therapies, heart failure remains a fatal disease. Myocardial infarction is the most common cause of heart failure, and only palliative measures are available to relieve symptoms and prolong the patient's life span. Because mammalian cardiomyocytes irreversibly exit the cell cycle at about the time of birth, the heart has traditionally been considered to lack any regenerative capacity. This paradigm, however, is currently shifting, and the cellular composition of the myocardium is being targeted by various regeneration strategies. Adult progenitor and stem cell treatment of diseased human myocardium has been carried out for more than 10 years (Menasche et al., 2001; Stamm et al., 2003), and it has become clear that, in humans, the regenerative capacity of hematopoietic stem cells and endothelial progenitor cells, despite potent proangiogenic effects, is limited (Stamm et al., 2009). More recently, mesenchymal stem cells (MSCs) and related cell types are being evaluated in preclinical models of heart disease as well as in clinical trials (see Published Clinical Trials, below). MSCs have the capacity to self-renew and to differentiate into lineages that normally originate from the embryonic mesenchyme (connective tissues, blood vessels, blood-related organs) (Caplan, 1991; Prockop, 1997; Pittenger et al., 1999). The current definition of MSCs includes plastic adherence in cell culture, specific surface antigen expression (CD105(+)/CD90(+)/CD73(+), CD34(-)/CD45(-)/CD11b(-) or CD14(-)/CD19(-) or CD79α(-)/HLA-DR1(-)), and multilineage in vitro differentiation potential (osteogenic, chondrogenic, and adipogenic) (Dominici et al., 2006 ). If those criteria are not met completely, the term "mesenchymal stromal cells" should be used for marrow-derived adherent cells, or other terms for MSC-like cells of different origin. For the purpose of this review, MSCs and related cells are discussed in general, and cell type-specific properties are indicated when appropriate. We first summarize the preclinical data on MSCs in models of heart disease, and then appraise the clinical experience with MSCs for cardiac cell therapy.

PMID: 21062128 [PubMed - indexed for MEDLINE]


CXCR4 signaling regulates metastasis of chemoresistant melanoma cells by a lymphatic metastatic niche.
Source:  NIH.gov
Monday, 29 November 1999 17:00

CXCR4 signaling regulates metastasis of chemoresistant melanoma cells by a lymphatic metastatic niche.

Cancer Res. 2010 Dec 15;70(24):10411-21

Authors: Kim M, Koh YJ, Kim KE, Koh BI, Nam DH, Alitalo K, Kim I, Koh GY

Abstract
Highly metastatic and chemotherapy-resistant properties of malignant melanomas stand as challenging barriers to successful treatment; yet, the mechanisms responsible for their aggressive characteristics are not fully defined. We show that a distinct population expressing CD133 (Prominin-1), which is highly enriched after administration of a chemotherapeutic drug, dacarbazine, has enhanced metastatic potential in vivo. CD133(+) tumor cells are located close to tumor-associated lymphatic vessels in metastatic organs such as the regional lymph nodes and lung. Lymphatic endothelial cells promote the migratory activity of a CD133(+) subset to target organs and regulation of lymphatic growth efficiently modulates the metastasis of CD133(+) tumor cells. We found that lymphatic vessels in metastatic tissues stimulate chemokine receptor 4 (CXCR4)(+)/CD133(+) cell metastasis to target organs by secretion of stromal cell-derived factor-1 (SDF-1). The CXCR4(+)/CD133(+) cells exhibited higher metastatic activity compared with CXCR4(-)/CD133(+) cells and, importantly, blockade of CXCR4 coupled with dacarbazine efficiently inhibited both tumor growth and metastasis; dacarbazine alone could not attenuate tumor metastasis. The current study demonstrates a previously unidentified role of the lymphatic microenvironment in facilitating metastasis of chemoresistant melanoma cells via a specific chemotactic axis, SDF-1/CXCR4. Our findings suggest that targeting the SDF-1/CXCR4 axis in addition to dacarbazine treatment could therapeutically block chemoresistant CD133(+) cell metastasis toward a lymphatic metastatic niche.

PMID: 21056990 [PubMed - indexed for MEDLINE]


Paracrine effects of mesenchymal stem cells in cisplatin-induced renal injury require heme oxygenase-1.
Source:  NIH.gov
Monday, 29 November 1999 17:00

Paracrine effects of mesenchymal stem cells in cisplatin-induced renal injury require heme oxygenase-1.

Am J Physiol Renal Physiol. 2011 Jan;300(1):F254-62

Authors: Zarjou A, Kim J, Traylor AM, Sanders PW, Balla J, Agarwal A, Curtis LM

Abstract
Multipotent mesenchymal stem cells (MSC) have become a popular and promising therapeutic approach in many clinical conditions. MSC are beneficial in animal models of acute kidney injury (AKI), by mediating differentiation-independent paracrine properties, and have prompted ongoing clinical trials to evaluate the safety and efficacy of MSC. Heme oxygenase-1 (HO-1) is induced in response to stress including AKI and has important anti-apoptotic, anti-inflammatory, and proangiogenic properties in these settings. We therefore examined whether HO-1 plays a role in the beneficial effects of MSC in AKI. We isolated MSC from bone marrow of age-matched HO-1+/+ and HO-1-/- mice. Our studies indicate that while differentiation of MSC into osteo- and adipocytic lineages did not differ between cells isolated from HO-1+/+ and HO-1-/- mice, MSC from HO-1-/- mice had significantly lower angiogenic potential. Moreover, HO-1-/- MSC demonstrated reduced expression and secretion of several important growth and proangiogenic factors (stromal cell-derived factor-1, vascular endothelial growth factor-A, and hepatocyte growth factor) compared with MSC derived from HO-1+/+ mice. In addition, conditioned medium of HO-1+/+ MSC rescued functional and morphological changes associated with cisplatin-induced AKI, while the HO-1-/--conditioned medium was ineffectual. Our studies indicate that HO-1 plays an important role in MSC-mediated protection. The results expand understanding of the renoprotective effects of MSC and may provide novel strategies to better utilize MSC in various disease models.

PMID: 21048024 [PubMed - indexed for MEDLINE]


[Experimental study of the effect of adipose stromal vascular fraction cells on the survival rate of fat transplantation].
Source:  NIH.gov
Monday, 29 November 1999 17:00

[Experimental study of the effect of adipose stromal vascular fraction cells on the survival rate of fat transplantation].

Zhonghua Zheng Xing Wai Ke Za Zhi. 2010 Jul;26(4):289-94

Authors: Fu BC, Gao JH, Lu F, Li J

Abstract
OBJECTIVE: To investigate the effect of adipose stromal vascular fraction cells (SVFs) on the survival rate of fat transplantation.
METHODS: 0. 5 ml autologous fat tissue was mixed with: 1) Di-labeled autologous SVFs ( Group A); 2) Di-labeled autologous adipose-derived stem cells (ASCs) (Group B); 3)Complete DMEM (Group C). And then the mixture was injected randomly under the back skin of 14 rabbits. The transplanted fat tissue in three groups was harvested at 6 months after implantation. Wet weight of fat grafts was measured for macroscopic aspects. After HE staining, blood vessel density, viable adipocytes and fibrous proliferation were counted respectively for histological evaluation. Trace of DiI-labeled ASCs in vivo was detected by fluorescent microscope.
RESULTS: The wet weight of fat grafts in group A (291.0 +/- 72.1) mg and group B (269.3 +/- 67.3) mg was significantly higher than that in group C (177.8 +/- 60.0) mg, but the difference between Group A and Group B was not significant. Histological analysis revealed that the fat grafts in group A and B was consisted predominantly of adipose tissue with less fat necrosis and fibrosis, compared with the fat grafts in group C. The grafts in both group A and B had significantly higher capillary density than those in the control group. Part of vascular endothelial cells were observed to origin from ectogenic DiI-labeled SVFs and ASCs.
CONCLUSIONS: The autologous isolated SVFs has a similar effect as autologous cultured ASCs to improve the survival rate of fat transplantation. And the former is more practical and safe, indicating a wide clinical application in the future.

PMID: 21046778 [PubMed - indexed for MEDLINE]


Mesenchymal stem cells transduced by stromal cell-derived factor-1α augment ischemic free flaps' survival.
Source:  NIH.gov
Monday, 29 November 1999 17:00

Mesenchymal stem cells transduced by stromal cell-derived factor-1α augment ischemic free flaps' survival.

Ann Plast Surg. 2011 Jan;66(1):92-7

Authors: Zhang FG, Yao Y, Feng Y, Hua CG, Tang XF

Abstract
Mesenchymal stem cells (MSCs) have the potential for differentiating into vascular endothelial cells. Stromal cell-derived factor-1α (SDF-1α) plays an important role in neovascularization of ischemic flaps. The authors evaluated the feasibility of applying MSCs transduced by SDF-1α gene to the treatment of early and partial ischemic free flaps survival. MSCs were isolated from Lewis rats and cultured in vitro. Recombinant adenovirus encoding SDF-1α gene (Ad-SDF-1α) was transduced into the MSCs. Lewis rats that underwent epigastric free flaps based on medial and lateral branches of superficial inferior epigastric vessels and femoral vessels were equally randomized into 4 groups, and injected with Ad-SDF-1α-transduced MSCs, MSCs, Ad-SDF-1α, and normal saline, respectively. Gene transduction, flaps survival, neovascularization, and expression level of SDF-1a protein were detected. The results showed that Ad-SDF-1α-transduced MSCs expressed higher SDF-1α both in vitro and in vivo, yielded more survival area, and resulted in higher neovascularization than any other groups. Interestingly, the necrotic sites of all free flaps were in the proximal end rather than in the distal end. In conclusion, Ad-SDF-1α-transduced MSCs can increase neovascularization of early and partial ischemic free flaps and augment the surviving areas.

PMID: 21042172 [PubMed - indexed for MEDLINE]


Effect of endothelial progenitor cells in neovascularization and their application in tumor therapy.
Source:  NIH.gov
Monday, 29 November 1999 17:00

Effect of endothelial progenitor cells in neovascularization and their application in tumor therapy.

Chin Med J (Engl). 2010 Sep;123(17):2454-60

Authors: Dong F, Ha XQ

Abstract
OBJECTIVE: To review the effect of endothelial progenitor cells in neovascularization as well as their application to the therapy of tumors.
DATA SOURCES: The data used in this review were mainly from PubMed for relevant English language articles published from 1997 to 2009. The search term was "endothelial progenitor cells".
STUDY SELECTION: Articles regarding the role of endothelial progenitor cells in neovascularization and their application to the therapy of tumors were selected.
RESULTS: Endothelial progenitor cells isolated from bone marrow, umbilical cord blood and peripheral blood can proliferate, mobilize and differentiate into mature endothelial cells. Experiments suggest endothelial progenitor cells take part in forming the tumor vascular through a variety of mechanisms related to vascular endothelial growth factor, matrix metalloproteinases, chemokine stromal cell-derived factor 1 and its receptor C-X-C receptor-4, erythropoietin, Notch signal pathway and so on. Evidence demonstrates that the number and function change of endothelial progenitor cells in peripheral blood can be used as a biomarker of the response of cancer patients to anti-tumor therapy and predict the prognosis and recurrence. In addition, irradiation temporarily increased endothelial cells number and decreased the endothelial progenitor cell counts in animal models. Meanwhile, in preclinical experiments, therapeutic gene-modified endothelial progenitor cells have been approved to attenuate tumor growth and offer a novel strategy for cell therapy and gene therapy of cancer.
CONCLUSIONS: Endothelial progenitor cells play a particular role in neovascularization and have attractively potential prognostic and therapeutic applications to malignant tumors. However, a series of problems, such as the definitive biomarkers of endothelial progenitor cells, their interrelationship with radiotherapy and their application in cell therapy and gene therapy of tumors, need further investigation.

PMID: 21034565 [PubMed - indexed for MEDLINE]


Very small embryonic-like stem cells in cardiovascular repair.
Source:  NIH.gov
Monday, 29 November 1999 17:00

Very small embryonic-like stem cells in cardiovascular repair.

Pharmacol Ther. 2011 Jan;129(1):21-8

Authors: Wojakowski W, Kucia M, Zuba-Surma E, Jadczyk T, Książek B, Ratajczak MZ, Tendera M

Abstract
Adult bone marrow (BM) harbors several small populations of cells which may contribute to cardiac and endothelial repair, such as endothelial progenitor cells (EPCs), mesenchymal stromal cells (MSCs) and very small embryonic-like cells (VSELs) expressing several markers of pluripotent stem cells (PSCs), such as Oct-4, Nanog and SSEA-1. Such cells were identified in mice bone marrow, peripheral blood and solid organs as well as in umbilical cord blood (UCB) and peripheral blood (PB) in humans. The adult BM-derived VSELs may undergo differentiation into cells derived for all three germ layers, including cardiomyocytes and vascular endothelial cells. VSELs can be isolated using a multiparameter live cell sorting technique with special gating strategy based on their small size, expression of stem cell markers (Sca-1 in mice, CXCR4 and CD133 in humans) and absence of hematopoietic lineage markers (CD45(-) Lin(-)). Experiments in murine models of myocardial infarction (MI) demonstrated population of VSELs expressed also early markers of cardiac and endothelial lineages (GATA-4, Nkx2.5/Csx, VE-cadherin, von Willebrand factor) which migrated to stromal-derived factor-1 (SDF-1) and other chemoattractant gradient and underwent rapid mobilization into peripheral blood in experimental MI mice models. Recently, we demonstrated the mobilization of VSELs expressing PSC, early cardiac and endothelial markers in patients with acute MI. In addition to BM, VSELs were also identified in several murine solid organs including the heart and brain, as well as in umbilical cord blood and peripheral blood in adult humans. We hypothesized that VSELs are quiescent progeny of epiblast-derived PSCs that are deposited during organogenesis in developing organs. In experimental MI intramyocardial injection of VSELs was more efficient than that of HSCs at improving left ventricular ejection fraction and attenuation of myocardial hypertrophy. VSELs can be useful in translational studies of cardiovascular repair.

PMID: 20971132 [PubMed - indexed for MEDLINE]


Coronary endothelial dysfunction in humans is associated with coronary retention of osteogenic endothelial progenitor cells.
Source:  NIH.gov
Monday, 29 November 1999 17:00

Coronary endothelial dysfunction in humans is associated with coronary retention of osteogenic endothelial progenitor cells.

Eur Heart J. 2010 Dec;31(23):2909-14

Authors: Gössl M, Mödder UI, Gulati R, Rihal CS, Prasad A, Loeffler D, Lerman LO, Khosla S, Lerman A

Abstract
AIMS: Endothelial progenitor cells (EPC) may participate in the repair of injured coronary endothelium. We have recently identified EPC co-expressing the osteoblastic marker osteocalcin [OCN (+) EPC] and found that their numbers are increased in patients with early and late coronary atherosclerosis. The current study was designed to test the hypothesis that early coronary atherosclerosis is associated with the retention of osteogenic EPC within the coronary circulation.
METHODS AND RESULTS: Blood samples were taken simultaneously from the proximal aorta and the coronary sinus from 31 patients undergoing invasive coronary endothelial function testing. Using flow cytometry, peripheral blood mononuclear cells were analysed for EPC markers (CD133, CD34, KDR) and OCN. The net gradient of EPC was calculated by multiplying the coronary blood flow by the arteriovenous EPC gradient (a negative net gradient indicating retention of EPC). Similarly, serum samples were analysed for stromal cell-derived factor-1 alpha (SDF-1 alpha) and interleukin-8 (IL-8) and their net production calculated. Compared with controls (n = 17) patients with endothelial dysfunction (ED, n = 14) had a significant net retention of CD34+/CD133-/KDR+/OCN+ EPC [118.38 (0.00, 267.04) vs. -112.03 (838.36, 0.00), P = 0.004]. The retention of OCN (+) EPC correlated with the degree of ED. Patients with ED also showed a net retention of CD34+/CD133-/KDR+ EPC (P = 0.010). Net production of IL-8 was positive in ED [1540.80 (-300.40, 21744.10)pg/mL] but negative in controls [-3428.50 (-11225.00, 647.48), P = 0.025].
CONCLUSION: Our study demonstrates that patients with early coronary atherosclerosis are characterized by retention of OCN (+) EPC within the coronary circulation, potentially leading to progressive coronary calcification rather than normal repair.

PMID: 20935001 [PubMed - indexed for MEDLINE]


Targeting of c-kit+ haematopoietic progenitor cells prevents hypoxic pulmonary hypertension.
Source:  NIH.gov
Monday, 29 November 1999 17:00

Targeting of c-kit+ haematopoietic progenitor cells prevents hypoxic pulmonary hypertension.

Eur Respir J. 2011 Jun;37(6):1392-9

Authors: Gambaryan N, Perros F, Montani D, Cohen-Kaminsky S, Mazmanian M, Renaud JF, Simonneau G, Lombet A, Humbert M

Abstract
Haematopoietic c-kit+ progenitor cells may contribute to pulmonary vascular remodelling and pulmonary hypertension (PH). Stromal derived factor-1 (SDF-1/CXCL12) and its receptors CXCR4 and CXCR7 have been shown to be critical for homing and mobilisation of haematopoietic c-kit+ progenitor cells in the perivascular niche. We administered AMD3100, a CXCR4 antagonist, and CCX771, a CXCR7 antagonist, to chronic hypoxia exposed mice in order to study the role of c-kit+ progenitor cells in PH. CXCL12, CXCR4 and CXCR7 protein expression, haemodynamic parameters, right ventricular mass, extent of vascular remodelling and perivascular progenitor cell accumulation were studied. Chronic hypoxia-exposed mice showed increased total lung tissue expression of CXCR4, CXCR7 and CXCL12 after development of PH. This was associated with significantly increased right ventricular systolic pressure and evidence of right ventricular hypertrophy, vascular remodelling and perivascular c-kit+/sca-1+ progenitor cell accumulation. CCX771 administration did not abrogate these effects. In contrast, administration of AMD3100, whether alone or combined with CCX771, prevented vascular remodelling, PH and perivascular accumulation of c-kit+/sca-1+ progenitor cells, with a synergistic effect of these agents. This study offers important pathophysiological insights into the role of haematopoietic c-kit+ progenitors in hypoxia-induced vascular remodelling and may have therapeutic implications for PH.

PMID: 20884740 [PubMed - indexed for MEDLINE]


Identification of E-selectin as a novel target for the regulation of postnatal neovascularization: implications for diabetic wound healing.
Source:  NIH.gov
Monday, 29 November 1999 17:00

Identification of E-selectin as a novel target for the regulation of postnatal neovascularization: implications for diabetic wound healing.

Ann Surg. 2010 Oct;252(4):625-34

Authors: Liu ZJ, Tian R, An W, Zhuge Y, Li Y, Shao H, Habib B, Livingstone AS, Velazquez OC

Abstract
OBJECTIVES: We previously reported that stromal cell-derived factor-1α (SDF-1α, a homing signal for recruiting endothelial progenitor cells (EPC) to areas of neovascularization), is down-regulated in diabetic wounds (Gallagher et al, J Clin Invest. 2007;117:1249-1259). We now investigate signals whereby mature endothelial cells (EC) and circulating EPC achieve SDF-1α-mediated EPC homing.
METHODS: SDF-1α in diabetic wounds were therapeutically increased by injection of SDF-1α-engineered bone marrow-derived fibroblasts versus control cells (N = 48 [20, non-obese diabetic (NOD)], [28, streptozotocin-C57]). Polymerase chain reaction-array gene expression differences were validated by Western blotting and immunohistochemistry. The role of adhesion molecule(s) in mediating SDF-1α-induced EPC homing, and wound healing was furthered studied using antagonists in vitro and in vivo.
RESULTS: Increasing wound SDF-1α via cell-based therapy promotes healing in diabetic mice (∼20% increase in healing rates by day 3, P = 0.006). SDF-1α increased EC-EPC adhesion and specifically upregulated E-selectin expression in human microvascular EC (2.3-fold increase, P < 0.01). This effect was also significant in blood vessels of the experimental mice and resulted in increased wound neovascularization. The regulatory effects of SDF-1α on EC-EPC adhesion and EPC homing were specifically mediated by E-selectin, as the application of E-selectin antagonists significantly inhibited SDF-1α-induced EC-EPC adhesion, EPC homing, wound neovascularization, and wound healing.
CONCLUSIONS: SDF-1α-engineered cell-based therapy promotes diabetic wound healing in mice by specifically upregulating E-selectin expression in mature EC leading to increase EC-EPC adhesion, EPC homing, and increased wound neovascularization. These findings provide novel insight into the signals underlying the biological effect of SDF-1α on EPC homing and point to E-selectin as a new potential target for therapeutic manipulation of EPC trafficking in diabetic wound healing.

PMID: 20881769 [PubMed - indexed for MEDLINE]


Evidence of endothelial progenitor cells in the human brain and spinal cord arteriovenous malformations.
Source:  NIH.gov
Monday, 29 November 1999 17:00

Evidence of endothelial progenitor cells in the human brain and spinal cord arteriovenous malformations.

Neurosurgery. 2010 Oct;67(4):1029-35

Authors: Gao P, Chen Y, Lawton MT, Barbaro NM, Yang GY, Su H, Ling F, Young WL

Abstract
BACKGROUND: Brain and spinal cord arteriovenous malformations (AVMs) are characterized by aberrant angiogenesis and vascular remodeling. Endothelial progenitor cells (EPCs) can be recruited by stromal cell-derived factor-1 (SDF-1), and participate in vascular remodeling in both physiological and pathological settings.
OBJECTIVE: To investigate whether there are increased EPC levels in the brain and spinal cord AVM nidus.
METHODS: Microsurgical specimens without endovascular embolization and radiosurgery from the brain (n = 12) and spinal cord (n = 5) AVMs were examined. Hemangioblastoma, meningioma, cerebral cortex obtained from epilepsy surgery, and the basilar artery from the autopsy were chosen for control comparisons. EPCs were identified as cells that were double-positive for the stem cell marker CD133 and the endothelial cell marker VEGFR-2 (vascular endothelial growth factor receptor-2 or KDR). In addition, SDF-1 was characterized by immunohistochemistry.
RESULTS: Both brain and spinal AVM tissues displayed more CD133-, SDF-1-, and CD68-positive signals than epilepsy and basilar artery control tissues. The level of EPCs was increased in the brain and spinal cord AVM nidus, mainly at the edge of the vessel wall. The expression of SDF-1 was colocalized with CD31-positive and α-smooth muscle cells, and was predominantly found within the vessel wall.
CONCLUSION: Our data demonstrate that EPCs are present in the nidus of the brain and spinal cord AVMs, which may mediate pathological vascular remodeling and impact the clinical course of AVMs.

PMID: 20881566 [PubMed - indexed for MEDLINE]


Stromal cell-derived factor-1alpha activation of tissue-engineered endothelial progenitor cell matrix enhances ventricular function after myocardial infarction by inducing neovasculogenesis.
Source:  NIH.gov
Monday, 29 November 1999 17:00

Stromal cell-derived factor-1alpha activation of tissue-engineered endothelial progenitor cell matrix enhances ventricular function after myocardial infarction by inducing neovasculogenesis.

Circulation. 2010 Sep 14;122(11 Suppl):S107-17

Authors: Frederick JR, Fitzpatrick JR, McCormick RC, Harris DA, Kim AY, Muenzer JR, Marotta N, Smith MJ, Cohen JE, Hiesinger W, Atluri P, Woo YJ

Abstract
BACKGROUND: Myocardial ischemia causes cardiomyocyte death, adverse ventricular remodeling, and ventricular dysfunction. Endothelial progenitor cells (EPCs) have been shown to ameliorate this process, particularly when activated with stromal cell-derived factor-1α (SDF), known to be the most potent EPC chemokine. We hypothesized that implantation of a tissue-engineered extracellular matrix (ECM) scaffold seeded with EPCs primed with SDF could induce borderzone neovasculogenesis, prevent adverse geometric remodeling, and preserve ventricular function after myocardial infarction.
METHODS AND RESULTS: Lewis rats (n=82) underwent left anterior descending artery ligation to induce myocardial infarction. EPCs were isolated, characterized, and cultured on a vitronectin/collagen scaffold and primed with SDF to generate the activated EPC matrix (EPCM). EPCM was sutured to the anterolateral left ventricular wall, which included the region of ischemia. Control animals received sutures but no EPCM. Additional groups underwent application of the ECM alone, ECM primed with SDF (ECM+SDF), and ECM seeded with EPCs but not primed with SDF (ECM+SDF). At 4 weeks, borderzone myocardial tissue demonstrated increased levels of vascular endothelial growth factor in the EPCM group. When compared to controls, Vessel density as assessed by immunohistochemical microscopy was significantly increased in the EPCM group (4.1 versus 6.2 vessels/high-powered field; P<0.001), and microvascular perfusion measured by lectin microangiography was enhanced 4-fold (0.7% versus 2.7% vessel volume/section volume; P=0.04). Comparisons to additional groups also showed a significantly improved vasculogenic response in the EPCM group. Ventricular geometry and scar fraction assessed by digital planimetric analysis of sectioned hearts exhibited significantly preserved left ventricular internal diameter (9.7 mm versus 8.6 mm; P=0.005) and decreased infarct scar formation expressed as percent of total section area (16% versus 7%; P=0.002) when compared with all other groups. In addition, EPCM animals showed a significant preservation of function as measured by echocardiography, pressure-volume conductance, and Doppler flow.
CONCLUSIONS: Extracellular matrix seeded with EPCs primed with SDF induces borderzone neovasculogenesis, attenuates adverse ventricular remodeling, and preserves ventricular function after myocardial infarction.

PMID: 20837901 [PubMed - indexed for MEDLINE]


Isogenic venous graft supported with bone marrow stromal cells as a natural conduit for bridging a 20 mm nerve gap.
Source:  NIH.gov
Monday, 29 November 1999 17:00

Isogenic venous graft supported with bone marrow stromal cells as a natural conduit for bridging a 20 mm nerve gap.

Microsurgery. 2010 Nov;30(8):639-45

Authors: Nijhuis TH, Brzezicki G, Klimczak A, Siemionow M

Abstract
In this study, we introduce a technique for bridging large neural gaps, using an isogenic vein graft supported with isogenic bone marrow stromal cells (BMSC). In three groups a nerve defect of 20 mm was bridged with a vein graft. Our first experimental group comprized an empty venous graft, in group II the venous nerve graft was filled with saline where as in group III the venous nerve graft was filled with BMSC. The animals were tested for functional recovery up to 3 months post repair. Our results show that the BMSC filled venous graft resulted in significantly better regeneration of the nerve defect compared to controls, as confirmed by the functional recovery measured by somatosensory evoked potentials, toe spread, pin prick, and gastrocnemius muscle index. Conclusively, the results confirm that the vein graft supported with BMSC is associated with better functional nerve regeneration.

PMID: 20842703 [PubMed - indexed for MEDLINE]


Zinc supplementation results in improved therapeutic potential of bone marrow-derived mesenchymal stromal cells in a mouse ischemic limb model.
Source:  NIH.gov
Monday, 29 November 1999 17:00

Zinc supplementation results in improved therapeutic potential of bone marrow-derived mesenchymal stromal cells in a mouse ischemic limb model.

Cytotherapy. 2011 Feb;13(2):156-64

Authors: Zhang D, Li Y, Zhu T, Zhang F, Yang Z, Miao D

Abstract
BACKGROUND AIMS: We wanted to determine whether zinc supplementation can inhibit bone marrow-derived mesenchymal stromal cell (MSC) apoptosis and enhance their tissue regenerative potential a in mouse ischemic hindlimb model.
METHODS: Rat bone marrow cells were cultured and the resulting MSC were passaged for 3-7 generations. The proliferation and apoptosis of MSC was examined by 3-[4,5-dimethyl-2-thiazol-2-yl]-2,5-diphenyltetrazolium bromide (MTT) assay and flow cytometry analysis. The activation of protein kinases B (Akt) was determined by Western blots. Vascular endothelial growth factor (VEGF) levels were examined by enzyme-linked immunosorbent assay. The mouse hindlimb ischemic model was established by ligating the right femoral artery. Mice received MSC, zinc-treated MSC or vehicle. The blood flow was assessed by laser Doppler imaging. The survival rate of donor cells was quantified by real-time polymerase chain reaction for the sex-determining region of the Y-chromosome (Sry). Angiogenesis was assessed by histochemical staining and immunofluoresence staining.
RESULTS: Supplementation with physiologic amounts of zinc caused a marked attenuation of cell apoptosis, enhanced cell viabilities, increased VEGF release and up-regulated Akt activation. Zinc-treated MSC delivered into ischemic hindlimbs resulted in significant improvements in limb blood perfusion by increased implanted MSC survival and stimulated angiogenesis.
CONCLUSIONS: This study demonstrates the potential of zinc supplement to enhance survival of engrafted MSC and ameliorate their tissue regenerative potential in a mouse ischemic hindlimb model.

PMID: 20839997 [PubMed - indexed for MEDLINE]


Successful bone marrow transplantation reveals the lack of endothelial progenitor cells mobilization in a patient with critical limb ischemia: a case report.
Source:  NIH.gov
Monday, 29 November 1999 17:00

Successful bone marrow transplantation reveals the lack of endothelial progenitor cells mobilization in a patient with critical limb ischemia: a case report.

Transplant Proc. 2010 Sep;42(7):2816-20

Authors: Cobellis G, Botti C, Taddeo A, Silvestroni A, Lillo S, Da Ponte A, Villa ML, Sica V, Della Bella S

Abstract
Restoring blood flow to ischemic tissue is a prerequisite for treatment of ischemic diseases. Cell-based therapy based on bone marrow transplantation is a promising option for patients with critical limb ischemia (CLI). The efficacy of cell therapies to augment neovascularization seems to involve endothelial progenitor cells (EPCs); however, the mechanisms underlying the efficacy have not been fully elucidated. Herein we have described the case of a young patient with severe CLI, who experienced a 24-month beneficial clinical response to autologous bone marrow transplantation. The exceptional amelioration enabled him to perform standardized maximal treadmill exercise test that demonstrated lack of exercise-induced EPC mobilization, despite adequate stromal-derived factor 1 and vascular endothelial growth factor responses. Therefore, tissue ischemia is not sufficient to promote the recruitment of EPCs that have been demonstrated to be involved in the recovery from ischemia. The local implantation of marrow-derived elements may provide cells and/or trophic factors, which have the capacity to augment angiogenesis, opening new approaches to the etiopathogenesis of the disease.

PMID: 20832596 [PubMed - indexed for MEDLINE]


Prolonged hypoxic culture and trypsinization increase the pro-angiogenic potential of human adipose tissue-derived stem cells.
Source:  NIH.gov
Monday, 29 November 1999 17:00

Prolonged hypoxic culture and trypsinization increase the pro-angiogenic potential of human adipose tissue-derived stem cells.

Cytotherapy. 2011 Mar;13(3):318-28

Authors: Rasmussen JG, Frøbert O, Pilgaard L, Kastrup J, Simonsen U, Zachar V, Fink T

Abstract
BACKGROUND AIMS: Transplantation of mesenchymal stromal cells (MSC), including adipose tissue-derived stem cells (ASC), is a promising option in the treatment of vascular disease. Short-term hypoxic culture of MSC augments secretion of anti-apoptotic and angiogenic cytokines. We hypothesized that prolonged hypoxic (1% and 5% oxygen) culture and trypsinization would augment ASC expression of anti-apoptotic and angiogenic cytokines and increase the angiogenic potential of ASC-conditioned media.
METHODS: The effects of prolonged hypoxic culture on growth and pro-angiogenic properties were investigated using human ASC cultured at 1%, 5% and 21% oxygen. The effect of trypsinization on the expression of pro-angiogenic genes was also determined.
RESULTS: Trypsinization induced up-regulation of the vascular endothelial growth factor (VEGF) and insulin-like growth factor-1 (IGF-1) genes independent of oxygen concentration. The expression of VEGF and IGF-1 was up-regulated in ASC cultured at 1% oxygen for 13 days compared with 4 days. The VEGF concentration in ASC-conditioned media was higher after prolonged hypoxic culture compared with short-term culture, while the IGF-1 and chemokine (CXC motif) ligand 12 (CXCL12) concentrations were unchanged. The VEGF receptor blocker SU5416 abolished angiogenesis in a cultured rat aortic ring model. Media from cells exposed to hypoxia increased angiogenesis, an effect that was dependent on factors other than just the VEGF concentration in the added media.
CONCLUSIONS: Optimization of the angiogenic potential of stem cell-based therapy in the treatment of vascular disease is important. We have demonstrated that prolonged hypoxic culture and trypsinization augment the therapeutic angiogenic potential of ASC.

PMID: 20795759 [PubMed - indexed for MEDLINE]


Expression of cytokines in rat brain with focal cerebral ischemia after grafting with bone marrow stromal cells and endothelial progenitor cells.
Source:  NIH.gov
Monday, 29 November 1999 17:00

Expression of cytokines in rat brain with focal cerebral ischemia after grafting with bone marrow stromal cells and endothelial progenitor cells.

Cytotherapy. 2011 Jan;13(1):46-53

Authors: He XY, Chen ZZ, Cai YQ, Xu G, Shang JH, Kou SB, Li M, Zhang HT, Duan CZ, Zhang SZ, Ke YQ, Zeng YJ, Xu RX, Jiang XD

Abstract
BACKGROUND AIMS: This study aimed to observe nine factors expressed in rat ischemic brain after transplantation of bone marrow stromal cells (BMSC) and/or endothelial progenitor cells (EPC). These factors were vascular endothelial growth factor (VEGF), stromal cell-derived factor-1 (SDF-1), basic fibroblast growth factor (bFGF), insulin-like growth factor (IGF-l), transforming growth factor-β (TGF-β), platelet-derived growth factor-BB (PDGF-BB), brain-derived neurotrophic factor (BDNF), glial cell line-derived neurotrophic factor (GDNF) and nerve growth factor (NGF).
METHODS: Adult Wistar rats were divided randomly into four groups: a vehicle group, BMSC group, EPC group and BMSC combined with EPC group. The rats were subjected to middle cerebral artery occlusion (MCAO) then implanted intravenously with 3 × 10(6) BMSC, EPC, BMSC/EPC or phosphate-buffered saline (PBS) 24 h after MCAO. Neurologic functional deficits were measured on days 1, 7, 14, 28 after transplantation. On day 7 after transplantation, quantitative reverse transcription (qRT)-polymerase chain reaction (PCR) and Western blot were employed to detect the expression of VEGF, SDF-1, bFGF, IGF-l, TGF-β, PDGF-BB, BDNF, GDNF and NGF.
RESULTS: The neurologic evaluation found that the neurologic severity scores were no different between the four groups on day 1, and the scores of rats in the BMSC/EPC group were significantly lower than those of rats in the other groups on days 7, 14 and 28 after transplantation. The expressions of bFGF, VEGF and BNDF were significantly higher in the BMSC/EPC group compared with the other groups.
CONCLUSIONS: The intravenous transplantation of BMSC combined with EPC could promote the functional rehabilitation of rats with focal cerebral ischemia, and the mechanism may be related to the enhanced expression of factors.

PMID: 20735164 [PubMed - indexed for MEDLINE]


[Ultrastructural changes of vascular endothelium in patients with chronic ischemia of the extremities after conduction of multipotent stromal cells from adipose tissue transplantation].
Source:  NIH.gov
Monday, 29 November 1999 17:00

[Ultrastructural changes of vascular endothelium in patients with chronic ischemia of the extremities after conduction of multipotent stromal cells from adipose tissue transplantation].

Klin Khir. 2010 Jun;(6):50-3

Authors: Poliachenko IuV, Driuk MF, Dombrovs'kyĭ DB

Abstract
Taking into account the impossibility of performance in some situations of reconstructive operative interventions on arteries, it is necessary to look for new methods of indirect revascularization for the extremities ischemia. Adipose tissue constitutes an accessible and sufficient source of multipotent stromal cells (MSC). Experimental investigations were made in a frame of preclinical trial on laboratory animals with the extremity ischemia simulation, and there was proved the essential stimulation of angiogenesis processes after transplantation performance of stromal-vascular fraction of adipose tissue. The work objective was to study the influence of own adipose tissue MSC transplantation on vascular endothelium changes in patients, suffering chronic ischemia of the extremities. Using electron microscopy method there was proved on microstructural level, that in clinical environment the patients, suffering chronic ischemia of the extremities of various etiology, gain undoubted effect of MSC autotransplantation performed with the objective to stimulate the processes of angiogenesis in the ischemic affection region.

PMID: 20734820 [PubMed - indexed for MEDLINE]


Efficacy assessment of interferon-alpha-engineered mesenchymal stromal cells in a mouse plasmacytoma model.
Source:  NIH.gov
Monday, 29 November 1999 17:00

Efficacy assessment of interferon-alpha-engineered mesenchymal stromal cells in a mouse plasmacytoma model.

Stem Cells Dev. 2011 Apr;20(4):709-19

Authors: Sartoris S, Mazzocco M, Tinelli M, Martini M, Mosna F, Lisi V, Indraccolo S, Moserle L, Cestari T, Riviera AP, Bifari F, Tridente G, Pizzolo G, Krampera M

Abstract
Bone marrow mesenchymal stromal cells (BM-MSCs) may survive and proliferate in the presence of cycling neoplastic cells. Exogenously administered MSCs are actively incorporated in the tumor as stromal fibroblasts, thus competing with the local mesenchymal cell precursors. For this reason, MSCs have been suggested as a suitable carrier for gene therapy strategies, as they can be genetically engineered with genes encoding for biologically active molecules that can inhibit tumor cell proliferation and enhance the antitumor immune response. We used BM-MSCs engineered with the murine interferon-alpha (IFN-α) gene (BM-MSCs/IFN-α) to assess in a mouse plasmacytoma model the efficacy of this approach toward neoplastic plasma cells. We found that IFN-α can be efficiently produced and delivered inside the tumor microenvironment. Subcutaneous multiple administration of BM-MSCs/IFN-α significantly hampered the tumor growth in vivo and prolonged the overall survival of mice. The antitumor effect was associated with enhanced apoptosis of tumor cells, reduction in microvessel density, and ischemic necrosis. By contrast, intravenous administration of BM-MSCs/IFN-α did not significantly modify the survival of mice, mainly as a consequence of an excessive entrapment of injected cells in the pulmonary vessels. In conclusion, BM-MSCs/IFN-α are effective in inhibiting neoplastic plasma cell growth; however, systemic administration of engineered MSCs needs to be improved to make this approach potentially suitable for the treatment of multiple myeloma.

PMID: 20695752 [PubMed - indexed for MEDLINE]


Activation of ephrin A proteins influences hematopoietic stem cell adhesion and trafficking patterns.
Source:  NIH.gov
Monday, 29 November 1999 17:00

Activation of ephrin A proteins influences hematopoietic stem cell adhesion and trafficking patterns.

Exp Hematol. 2010 Nov;38(11):1087-98

Authors: Ting MJ, Day BW, Spanevello MD, Boyd AW

Abstract
OBJECTIVE: To determine if Eph receptors and ephrins can modulate the homing of hematopoietic cells in a murine bone marrow transplantation model.
MATERIALS AND METHODS: EphA and ephrin A gene expression by mouse hematopoietic stem cells and the progenitor cell line FDCP-1 was determined by real-time reverse transcription polymerase chain reaction and flow cytometry. The effect of ephrin A activation on adhesion of hematopoietic progenitors was determined by in vitro adhesion assays in which cells were exposed to fibronectin or vascular cell adhesion molecule-1 (VCAM-1) and an increasing gradient of immobilized EphA3-Fc. Adhesion to fibronectin and VCAM-1 was further investigated using soluble preclustered EphA3-Fc. We used soluble unclustered EphA3-Fc as an antagonist to block endogenous EphA-ephrin A interactions in vivo. The effect of injecting soluble EphA3-Fc on the mobilization of hematopoietic progenitor cells was examined. We determined the effect on short-term homing by pretreating bone marrow cells with EphA3-Fc or the control IgG before infusion into lethally irradiated mice.
RESULTS: Preclustered and immobilized EphA3-Fc increased adhesion of progenitor cells and FDCP-1 to fibronectin and VCAM-1 (1.6- to 2-fold higher adhesion; p < 0.05) relative to control (0 μ/cm(2) EphA3-Fc extracellular molecule alone). Injection of the antagonist soluble EphA3-Fc increased progenitor cell and colony-forming unit-spleen cells in the peripheral blood (42% greater colony-forming unit in culture; p < 0.05, 3.8-fold higher colony-forming unit-spleen) relative to control.
CONCLUSION: Treating bone marrow cells with EphA3-Fc resulted in a reduction by 31% in donor stem cells homing to the bone marrow and accumulation of donor cells in recipient spleens (50% greater than control) and greater recovery of donor stem cells from the peripheral blood.

PMID: 20655977 [PubMed - indexed for MEDLINE]


Recruitment of myeloid but not endothelial precursor cells facilitates tumor regrowth after local irradiation.
Source:  NIH.gov
Monday, 29 November 1999 17:00

Recruitment of myeloid but not endothelial precursor cells facilitates tumor regrowth after local irradiation.

Cancer Res. 2010 Jul 15;70(14):5679-85

Authors: Kozin SV, Kamoun WS, Huang Y, Dawson MR, Jain RK, Duda DG

Abstract
Tumor neovascularization and growth might be promoted by the recruitment of bone marrow-derived cells (BMDC), which include endothelial precursor cells and "vascular modulatory" myelomonocytic (CD11b+) cells. BMDCs may also drive tumor regrowth after certain chemotherapeutic and vascular disruption treatments. In this study, we evaluated the role of BMDC recruitment in breast and lung carcinoma xenograft models after local irradiation (LI). We depleted the bone marrow by including whole-body irradiation (WBI) of 6 Gy as part of a total tumor dose of 21 Gy, and compared the growth delay with the one achieved after LI of 21 Gy. In both models, the inclusion of WBI induced longer tumor growth delays. Moreover, WBI increased lung tumor control probability by LI. Exogenous delivery of BMDCs from radiation-naïve donors partially abrogated the WBI effect. Myeloid BMDCs, primarily macrophages, rapidly accumulated in tumors after LI. Intratumoral expression of stromal-derived factor 1alpha (SDF-1alpha), a chemokine that promotes tissue retention of BMDCs, was noted 2 days after LI. Conversely, treatment with an inhibitor of SDF-1alpha receptor CXCR4 (AMD3100) with LI significantly delayed tumor regrowth. However, when administered starting from 5 days post-LI, AMD3100 treatment was ineffective. Lastly, with restorative bone marrow transplantation of Tie2-GFP-labeled BMDC population, we observed an increased number of monocytes but not endothelial precursor cells in tumors that recurred following LI. Our results suggest that an increase in intratumoral SDF-1alpha triggered by LI recruits myelomonocytes/macrophages which promotes tumor regrowth.

PMID: 20631066 [PubMed - indexed for MEDLINE]


Protein therapeutics for cardiac regeneration after myocardial infarction.
Source:  NIH.gov
Monday, 29 November 1999 17:00

Protein therapeutics for cardiac regeneration after myocardial infarction.

J Cardiovasc Transl Res. 2010 Oct;3(5):469-77

Authors: Segers VF, Lee RT

Abstract
Although most medicines have historically been small molecules, many newly approved drugs are derived from proteins. Protein therapies have been developed for treatment of diseases in almost every organ system, including the heart. Great excitement has now arisen in the field of regenerative medicine, particularly for cardiac regeneration after myocardial infarction. Every year, millions of people suffer from acute myocardial infarction, but the adult mammalian myocardium has limited regeneration potential. Regeneration of the heart after myocardium infarction is therefore an exciting target for protein therapeutics. In this review, we discuss different classes of proteins that have therapeutic potential to regenerate the heart after myocardial infarction. Protein candidates have been described that induce angiogenesis, including fibroblast growth factors and vascular endothelial growth factors, although thus far clinical development has been disappointing. Chemotactic factors that attract stem cells, e.g., hepatocyte growth factor and stromal cell-derived factor-1, may also be useful. Finally, neuregulins and periostin are proteins that induce cell-cycle reentry of cardiomyocytes, and growth factors like IGF-1 can induce growth and differentiation of stem cells. As our knowledge of the biology of regenerative processes and the role of specific proteins in these processes increases, the use of proteins as regenerative drugs could develop as a cardiac therapy.

PMID: 20607468 [PubMed - indexed for MEDLINE]


Adipose tissue as a stem cell source for musculoskeletal regeneration.
Source:  NIH.gov
Monday, 29 November 1999 17:00

Adipose tissue as a stem cell source for musculoskeletal regeneration.

Front Biosci (Schol Ed). 2011;3:69-81

Authors: Gimble JM, Grayson W, Guilak F, Lopez MJ, Vunjak-Novakovic G

Abstract
Adipose tissue is an abundant, easily accessible, and reproducible cell source for musculo-skeletal regenerative medicine applications. Initial derivation steps yield a heterogeneous population of cells of stromal vascular fraction (SVF) cells. Subsequent adherent selection of the SVF results in a relatively homogeneous population of adipose-derived stromal/stem cells (ASCs) capable of adipogenic, chondrogenic, myogenic, and osteogenic differentiation in vitro on scaffolds in bioreactors and in vivo in pre-clinical animal models. Unlike hematopoietic cells, ASCs do not elicit a robust lymphocyte reaction and instead release immunosuppressive factors, such as prostaglandin E2. These immunomodulatory features suggest that allogeneic and autologous ASCs will engraft successfully for tissue regeneration purposes. The differentiation and expansion potential of ASCs can be modified by growth factors, bio-inductive scaffolds, and bioreactors providing environmental control and biophysical stimulation. Gene therapy approaches using lentiviral transduction can be used to direct differentiation of ASCs to particular lineages. We discuss the utility of ASCs for musculo-skeletal tissue repair and some of the technologies that can be implemented to unlock the full regenerative potential of these highly valuable cells.

PMID: 21196358 [PubMed - indexed for MEDLINE]


Stem cell therapy for Idiopathic Pulmonary Fibrosis: A protocol proposal.
Source:  NIH.gov
Monday, 29 November 1999 17:00

Stem cell therapy for Idiopathic Pulmonary Fibrosis: A protocol proposal.

J Transl Med. 2011 Oct 21;9(1):182

Authors: Tzouvelekis A, Koliakos G, Ntolios P, Baira I, Bouros E, Oikonomou A, Zissimopoulos A, Kolios G, Kakagia D, Paspaliaris V, Kotsianidis I, Froudarakis M, Bouros D

Abstract
ABSTRACT: BACKGROUND: Idiopathic pulmonary fibrosis represents a lethal form of progressive fibrotic lung disorder with gradually increasing incidence worldwide. Despite intense research efforts its pathogenesis is still elusive and controversial reflecting in the current disappointing status regarding its treatment. Patients and Methods: We report the first protocol proposal of a prospective, unicentric, non-randomized, phase Ib clinical trial to study the safety and tolerability of the adipose-derived stem cells (ADSCs) stromal vascular fraction (SVF) as a therapeutic agent in IPF. After careful patient selection based on functional criteria (forced vital capacity-FVC>50%, diffuse lung capacity for carbon monoxide-DLCO>35% of the predicted values) all eligible subjects will be subjected to lipoaspiration resulting in the isolation of approximately 100- 500 gr of adipose tissue. After preparation, isolation and labelling ADSCs-SVF will be endobronchially infused to both lower lobes of the fibrotic lungs. Procedure will be repeated thrice at monthly intervals. Primary end-point represent safety and tolerability data, while exploratory secondary end-points include assessment of clinical functional and radiological status. Results: Preliminary results recently presented in the form of an abstract seem promising and tantalizing since there were no cases of clinically significant allergic reactions, infections, disease acute exacerbations or ectopic tissue formation. In addition 6 months follow-up data revealed a marginal improvement at 6-minute walking distance and forced vital capacity. CONCLUSIONS: Adipose tissue represents an abundant, safe, ethically uncontested and potentially beneficial source of stem cells for patients with IPF. Larger multicenter phase II and III placebo-controlled clinical trials are sorely needed in order to prove efficacy. However, pilot safety studies are of major importance and represent the first hamper that should be overcome to establish a rigid basis for larger clinical trials.

PMID: 22017817 [PubMed - as supplied by publisher]


Stem-cell therapy in an experimental model of pulmonary hypertension and right heart failure: Role of paracrine and neurohormonal milieu in the remodeling process.
Source:  NIH.gov
Monday, 29 November 1999 17:00

Stem-cell therapy in an experimental model of pulmonary hypertension and right heart failure: Role of paracrine and neurohormonal milieu in the remodeling process.

J Heart Lung Transplant. 2011 Nov;30(11):1281-93

Authors: Angelini A, Castellani C, Ravara B, Franzin C, Pozzobon M, Tavano R, Libera LD, Papini E, Vettor R, De Coppi P, Thiene G, Vescovo G

Abstract
BACKGROUND: In this study we investigated the effect of human amniotic fluid stem (hAFS) cells and rat adipose tissue stromal vascular fraction GFP-positive cell (rSVC-GFP) therapy and the contribution of the paracrine and neurohormonal milieu to cardiac and pulmonary vascular remodeling in a rat model of pulmonary hypertension (PH) and right heart failure (RHF).
METHODS: Sprague-Dawley rats were injected with monocrotaline (MCT). Four million hAFS or rSVC-GFP cells were injected via the tail vein 3 weeks after MCT. RHF was confirmed by RV hypertrophy/dilation and by brain natriuretic peptide (BNP) level. Cytokine profile was assessed by Multiplex array. Stem cell (SC) differentiation was studied by immunofluorescence.
RESULTS: MCT rats showed eccentric RV hypertrophy with increased RV dilation (measured as right ventricular mass/right ventricular volume [RVM/RVV]: MCT, 1.46 ± 0.12; control, 2.33 ± 0.24; p = 0.01), and increased RV hypertrophy (measured as LVM/RVM: MCT, 1.58 ± 0.06; control, 2.83 ± 0.1; p < 0.00001), increased BNP (MCT, 5.2 ± 1.2; control, 1.5 ± 0.1; p < 0.001) and both pro- and anti-inflammatory cytokines. SC produced a fall of BNP (hAFS, 2.1 ± 0.7; rSVC-GFP, 1.98 ± 1.3; p < 0.001) and pro-inflammatory cytokines. Positive RV remodeling with decreased RV dilation (RVM/RVV: hAFS, 1.87 ± 0.44; rSVC-GFP, 2.12 ± 0.24; p < 0.03 and p < 0.05 vs MCT) and regression of RV hypertrophy (LVM/RVM: hAFS, 2.06 ± 0.08; rSVC-GFP, 2.16 ± 0.08; p < 0.00001 vs MCT) was seen together with a decrease in medial wall thickness of pulmonary arterioles (hAFS, 35.33 ± 2.78%; rSVC-GFP, 37.15 ± 2.92%; p = 0.0001 vs MCT).
CONCLUSIONS: SC engrafted in the lung, heart and skeletal muscle modulated the pro- and anti-inflammatory cytokine milieu, and produced a positive neurohormonal response. This was accompanied by positive cardiac and pulmonary vascular remodeling, with formation mainly of new vascular cells.

PMID: 21989772 [PubMed - in process]


Bone Marrow Suppression by c-Kit Blockade Enhances Tumor Growth of Colorectal Metastases through the Action of Stromal Cell-Derived Factor-1.
Source:  NIH.gov
Monday, 29 November 1999 17:00

Bone Marrow Suppression by c-Kit Blockade Enhances Tumor Growth of Colorectal Metastases through the Action of Stromal Cell-Derived Factor-1.

J Oncol. 2012;2012:196957

Authors: Rupertus K, Haberl GC, Scheuer C, Menger MD, Schilling MK, Kollmar O

Abstract
Background. Mobilization of c-Kit(+) hematopoietic cells (HCs) contributes to tumor vascularization. Whereas survival and proliferation of HCs are regulated by binding of the stem cell factor to its receptor c-Kit, migration of HCs is directed by stromal cell-derived factor (SDF)-1. Therefore, targeting migration of HCs provides a promising new strategy of anti-tumor therapy. Methods. BALB/c mice (n = 16) were pretreated with an anti-c-Kit antibody followed by implantation of CT26.WT-GFP colorectal cancer cells into dorsal skinfold chambers. Animals (n = 8) additionally received a neutralizing anti-SDF-1 antibody. Animals (n = 8) treated with a control antibody served as controls. Investigations were performed using intravital fluorescence microscopy, immunohistochemistry, flow cytometry and western blot analysis. Results. Blockade of c-Kit significantly enhanced tumor cell engraftment compared to controls due to stimulation of tumor cell proliferation and invasion without markedly affecting tumor vascularization. C-Kit blockade significantly increased VEGF and CXCR4 expression within the growing tumors. Neutralization of SDF-1 completely antagonized this anti-c-Kit-associated tumor growth by suppression of tumor neovascularization, inhibition of tumor cell proliferation and reduction of muscular infiltration. Conclusion. Our study indicates that bone marrow suppression via anti-c-Kit pretreatment enhances tumor cell engraftment of colorectal metastases due to interaction with the SDF-1/CXCR4 pathway which is involved in HC-mediated tumor angiogenesis.

PMID: 21977032 [PubMed - in process]


Sunitinib-induced hyperammonemic encephalopathy in gastrointestinal stromal tumors.
Source:  NIH.gov
Monday, 29 November 1999 17:00

Sunitinib-induced hyperammonemic encephalopathy in gastrointestinal stromal tumors.

Ann Pharmacother. 2011 Oct;45(10):e56

Authors: Lee NR, Yhim HY, Yim CY, Kwak JY, Song EK

Abstract
OBJECTIVE: To report 2 cases of hyperammonemic encephalopathy induced by sunitinib in patients with metastatic gastrointestinal stromal tumor (GIST).
CASE SUMMARY: A 58-year-old man with imatinib-resistant metastatic GIST presented to the emergency department with confusion that developed 17 days after the initiation of sunitinib 50 mg/day. His serum ammonia level was markedly elevated (210 μg/dL). Sunitinib was discontinued, and an enema with lactulose was administered every hour. His neurologic status normalized within 24 hours and his serum ammonia level decreased to 64 μg/dL. A 68-year-old woman with imatinib-resistant metastatic GIST was admitted into the emergency department with confusion and irritability that developed 10 days after the start of sunitinib therapy. Her serum ammonia level was markedly elevated (389 μg/dL). Sunitinib was discontinued, and an enema with lactulose was administered every hour. Within 24 hours, her mental status was improved and her serum ammonia level was decreased to 116 μg/dL. Sunitinib was reintroduced, and the same symptoms occurred after day 7 of administration. Sunitinib was not prescribed afterward and the woman did not experience any further encephalopathic symptoms.
DISCUSSION: Sunitinib is a small molecule that inhibits multiple receptor tyrosine kinases such as stem cell factor receptor, vascular endothelial growth factor, and platelet-derived growth factor. It is used as second-line therapy for patients with imatinib-resistant GIST. Hyperammonemic encephalopathy is an uncommon fatal complication of chemotherapy. According to the Naranjo probability scale, sunitinib was a probable cause of hyperammonemic encephalopathy in the patients described here. Although the mechanism of hyperammonemia is unclear, hyperammonemic encephalopathy might be caused by a vascular disorder related to the antiangiogenic properties of sunitinib, and it has ethnic differences associated with genetic polymorphisms.
CONCLUSIONS: Sunitinib may induce hyperammonemic encephalopathy in some patients. Although further studies are warranted, clinicians should be aware of this severe adverse event when using sunitinib for treatment of GIST.

PMID: 21954449 [PubMed - in process]


Computational protein design to reengineer stromal cell-derived factor-1α generates an effective and translatable angiogenic polypeptide analog.
Source:  NIH.gov
Monday, 29 November 1999 17:00

Computational protein design to reengineer stromal cell-derived factor-1α generates an effective and translatable angiogenic polypeptide analog.

Circulation. 2011 Sep 13;124(11 Suppl):S18-26

Authors: Hiesinger W, Perez-Aguilar JM, Atluri P, Marotta NA, Frederick JR, Fitzpatrick JR, McCormick RC, Muenzer JR, Yang EC, Levit RD, Yuan LJ, Macarthur JW, Saven JG, Woo YJ

Abstract
BACKGROUND: Experimentally, exogenous administration of recombinant stromal cell-derived factor-1α (SDF) enhances neovasculogenesis and cardiac function after myocardial infarction. Smaller analogs of SDF may provide translational advantages including enhanced stability and function, ease of synthesis, lower cost, and potential modulated delivery via engineered biomaterials. In this study, computational protein design was used to create a more efficient evolution of the native SDF protein.
METHODS AND RESULTS: Protein structure modeling was used to engineer an SDF polypeptide analog (engineered SDF analog [ESA]) that splices the N-terminus (activation and binding) and C-terminus (extracellular stabilization) with a diproline segment designed to limit the conformational flexibility of the peptide backbone and retain the relative orientation of these segments observed in the native structure of SDF. Endothelial progenitor cells (EPCs) in ESA gradient, assayed by Boyden chamber, showed significantly increased migration compared with both SDF and control gradients. EPC receptor activation was evaluated by quantification of phosphorylated AKT, and cells treated with ESA yielded significantly greater phosphorylated AKT levels than SDF and control cells. Angiogenic growth factor assays revealed a distinct increase in angiopoietin-1 expression in the ESA- and SDF-treated hearts. In addition, CD-1 mice (n=30) underwent ligation of the left anterior descending coronary artery and peri-infarct intramyocardial injection of ESA, SDF-1α, or saline. At 2 weeks, echocardiography demonstrated a significant gain in ejection fraction, cardiac output, stroke volume, and fractional area change in mice treated with ESA compared with controls.
CONCLUSIONS: Compared with native SDF, a novel engineered SDF polypeptide analog (ESA) more efficiently induces EPC migration and improves post-myocardial infarction cardiac function and thus offers a more clinically translatable neovasculogenic therapy.

PMID: 21911811 [PubMed - indexed for MEDLINE]


Reduction of infarct size by intravenous injection of uncultured adipose derived stromal cells in a rat model is dependent on the time point of application.
Source:  NIH.gov
Monday, 29 November 1999 17:00

Reduction of infarct size by intravenous injection of uncultured adipose derived stromal cells in a rat model is dependent on the time point of application.

Stem Cell Res. 2011 Nov;7(3):219-29

Authors: van Dijk A, Naaijkens BA, Jurgens WJ, Nalliah K, Sairras S, van der Pijl RJ, Vo K, Vonk AB, van Rossum AC, Paulus WJ, van Milligen FJ, Niessen HW

Abstract
Stem cell therapy is a promising tool to improve outcome after acute myocardial infarction (AMI), but needs to be optimized since results from clinical applications remain ambiguous. A potent source of stem cells is the stromal vascular fraction of adipose tissue (SVF), which contains high numbers of adipose derived stem cells (ASC). We hypothesized that: 1) intravenous injection can be used to apply stem cells to the heart. 2) Uncultured SVF cells are easier and safer when cultured ASCs. 3) Transplantation after the acute inflammation period of AMI is favorable over early injection. For this, AMI was induced in rats by 40min of coronary occlusion. One or seven days after AMI, rats were intravenously injected with vehicle, 5×10(6) uncultured rat SVF cells or 1×10(6) rat ASCs. Rats were analyzed 35 days after AMI. Intravenous delivery of both fresh SVF cells and cultured ASCs 7 days after AMI significantly reduced infarct size compared to vehicle. Similar numbers of stem cells were found in the heart, after treatment with fresh SVF cells and cultured ASCs. Importantly, no adverse effects were found after injection of SVF cells. Using cultured ASCs, however, 3 animals had shortness of breath, and one animal died during injection. In contrast to application at 7 days post AMI, injection of SVF cells 1 day post AMI resulted in a small but non-significant infarct reduction (p=0.35). Taken together, intravenous injection of uncultured SVF cells subsequent to the acute inflammation period, is a promising stem cell therapy for AMI.

PMID: 21907165 [PubMed - in process]


Effects on proliferation and differentiation of multipotent bone marrow stromal cells engineered to express growth factors for combined cell and gene therapy.
Source:  NIH.gov
Monday, 29 November 1999 17:00

Effects on proliferation and differentiation of multipotent bone marrow stromal cells engineered to express growth factors for combined cell and gene therapy.

Stem Cells. 2011 Nov;29(11):1727-37

Authors: Fierro FA, Kalomoiris S, Sondergaard CS, Nolta JA

Abstract
A key mechanism for mesenchymal stem cells/bone marrow stromal cells (MSCs) to promote tissue repair is by secretion of soluble growth factors (GFs). Therefore, clinical application could be optimized by a combination of cell and gene therapies, where MSCs are genetically modified to express higher levels of a specific factor. However, it remains unknown how this overexpression may alter the fate of the MSCs. Here, we show effects of overexpressing the growth factors, such as basic fibroblast growth factor (bFGF), platelet derived growth factor B (PDGF-BB), transforming growth factor β(1) (TGF-β(1) ), and vascular endothelial growth factor (VEGF), in human bone marrow-derived MSCs. Ectopic expression of bFGF or PDGF-B lead to highly proliferating MSCs and lead to a robust increase in osteogenesis. In contrast, adipogenesis was strongly inhibited in MSCs overexpressing PDGF-B and only mildly affected in MSCs overexpressing bFGF. Overexpression of TGF-β(1) blocked both osteogenic and adipogenic differentiation while inducing the formation of stress fibers and increasing the expression of the smooth muscle marker calponin-1 and the chondrogenic marker collagen type II. In contrast, MSCs overexpressing VEGF did not vary from control MSCs in any parameters, likely due to the lack of VEGF receptor expression on MSCs. MSCs engineered to overexpress VEGF strongly induced the migration of endothelial cells and enhanced blood flow restoration in a xenograft model of hind limb ischemia. These data support the rationale for genetically modifying MSCs to enhance their therapeutically relevant trophic signals, when safety and efficacy can be demonstrated, and when it can be shown that there are no unwanted effects on their proliferation and differentiation. STEM CELLS 2011;29:1727-1737.

PMID: 21898687 [PubMed - in process]


The vascular niche: home for normal and malignant hematopoietic stem cells.
Source:  NIH.gov
Monday, 29 November 1999 17:00

The vascular niche: home for normal and malignant hematopoietic stem cells.

Leukemia. 2011 Sep 2;

Authors: Doan PL, Chute JP

Abstract
Hematopoietic stem cells (HSCs) are uniquely capable of self-renewal and provision of all of the mature elements of the blood and immune system throughout the lifetime of an individual. HSC self-renewal is regulated by both intrinsic mechanisms and extrinsic signals mediated via specialized microenvironments or 'niches' wherein HSCs reside. HSCs have been shown to reside in close association with bone marrow (BM) osteoblasts in the endosteal niche and also in proximity to BM sinusoidal vessels. An unresolved question surrounds whether the endosteal and vascular niches provide synchronous or redundant regulation of HSC fate or whether these niches provide wholly unique regulatory functions. Furthermore, while some aspects of the mechanisms through which osteoblasts regulate HSC fate have been defined, the mechanisms through which the vascular niche regulates HSC fate remain obscure. Here, we summarize the anatomic and functional basis supporting the concept of an HSC vascular niche as well as the precise function of endothelial cells, perivascular cells and stromal cells within the niche in regulating HSC fate. Lastly, we will highlight the role of the vascular niche in regulating leukemic stem cell fate in vivo.Leukemia advance online publication, 2 September 2011; doi:10.1038/leu.2011.236.

PMID: 21886170 [PubMed - as supplied by publisher]


Pericytes: developmental, physiological, and pathological perspectives, problems, and promises.
Source:  NIH.gov
Monday, 29 November 1999 17:00

Pericytes: developmental, physiological, and pathological perspectives, problems, and promises.

Dev Cell. 2011 Aug 16;21(2):193-215

Authors: Armulik A, Genové G, Betsholtz C

Abstract
Pericytes, the mural cells of blood microvessels, have recently come into focus as regulators of vascular morphogenesis and function during development, cardiovascular homeostasis, and disease. Pericytes are implicated in the development of diabetic retinopathy and tissue fibrosis, and they are potential stromal targets for cancer therapy. Some pericytes are probably mesenchymal stem or progenitor cells, which give rise to adipocytes, cartilage, bone, and muscle. However, there is still confusion about the identity, ontogeny, and progeny of pericytes. Here, we review the history of these investigations, indicate emerging concepts, and point out problems and promise in the field of pericyte biology.

PMID: 21839917 [PubMed - indexed for MEDLINE]


Involvement of ERK phosphorylation in the prevention of ischemia-induced ovarian follicular depletion by stem cells.
Source:  NIH.gov
Monday, 29 November 1999 17:00

Involvement of ERK phosphorylation in the prevention of ischemia-induced ovarian follicular depletion by stem cells.

Chin J Physiol. 2010 Jun 30;53(3):167-77

Authors: Tsai SC, Chen CP, Su TH, Kau MM, Lu CC

Abstract
Ovarian failure is commonly caused by aging, autoimmune disease, menopause and cancer therapy. We used an ischemic model in the ovary to test the hypothesis that stem cells are helpful for ovarian regeneration after injury. Three treatment regimes were employed: sham-operated control, ligation plus PBS, and ligation plus immortalized human bone marrow stromal cells (stem cells) groups. After ligation-induced ischemia, stem cells or PBS were injected into rat ovaries. Then, pregnant mare serum gonadotropin was given intra-peritoneally to initiate folliculogenesis. The animals were then sacrificed. The ovary gland was weighed, and ovarian folliculogenesis, stem cell differentiation and vascular neogenesis were evaluated. In order to study improvement of folliculogenesis after ovarian ischemia, steroidogenic acute regulatory protein (StAR), p44/p42 MAPK (T-ERK1/2), and phospho-p44/ p42 MAPK (P-ERK1/2) expression were specifically evaluated. Results indicated that ovarian size was smaller and that the rate of folliculogenesis was lower in ovarian ischemic-reperfusion animals, but both recovered after stem cell treatment. The stem cells migrated into the ovary and differentiated into theca cells, granulosa cells, corona radiata cells and vascular endothelial cells. In addition, von Willebrand factor (vWF) expression was increased; 17beta-estradiol (E2), progesterone (P4), P-ERK1/2 and StAR protein expression was recovered by stem cells treatment in the ischemic ovaries. The serum LH was significantly increased in ovaries of ischemia-reperfusion animals, but the stem cell treatment restored the effects. These results suggest that stem cells might be helpful for ovarian regeneration after injuries by promoting vascular neogenesis and steroidogenesis through the MAPK pathway.

PMID: 21793325 [PubMed - indexed for MEDLINE]


Amnion-Derived Mesenchymal Stromal Cells Show Angiogenic Properties but Resist Differentiation into Mature Endothelial Cells.
Source:  NIH.gov
Monday, 29 November 1999 17:00

Amnion-Derived Mesenchymal Stromal Cells Show Angiogenic Properties but Resist Differentiation into Mature Endothelial Cells.

Stem Cells Dev. 2011 Sep 6;

Authors: König J, Huppertz B, Desoye G, Parolini O, Fröhlich JD, Weiss G, Dohr G, Sedlmayr P, Lang I

Abstract
Mesenchymal stromal cells derived from the human amnion (hAMSC) currently play an important role in stem cell research, as they are multipotent cells that can be isolated using noninvasive methods and are immunologically tolerated in vivo. The objective of this study was to evaluate their endothelial differentiation potential with regard to a possible therapeutic use in vascular diseases. hAMSC were isolated from human term placentas and cultured in Dulbecco's modified Eagle's medium (DMEM) (non-induced hAMSC) or endothelial growth medium (EGM-2) (induced hAMSC). Induced hAMSC changed their fibroblast-like toward an endothelial-like morphology, and were able to take up acetylated low-density lipoprotein and form endothelial-like networks in the Matrigel assay. However, they did not express the mature endothelial cell markers von Willebrand factor and vascular endothelial-cadherin. Gene expression analysis revealed that induced hAMSC significantly downregulated pro-angiogenic genes such as tenascin C, Tie-2, vascular endothelial growth factor A (VEGF-A), CD146, and fibroblast growth factor 2 (FGF-2), whereas they significantly upregulated anti-angiogenic genes such as serpinF1, sprouty1, and angioarrestin. Analysis of protein expression confirmed the downregulation of FGF-2 and Tie-2 (27%±8% and 13%±1% of non-induced cells, respectively) and upregulation of the anti-angiogenic protein endostatin (226%±4%). Conditioned media collected from hAMSC enhanced viability of endothelial cells and had a stabilizing effect on endothelial network formation as shown by lactate dehydrogenase and Matrigel assay, respectively. In summary, endothelial induced hAMSC acquired some angiogenic properties but resisted undergoing a complete differentiation into mature endothelial cells by upregulation of anti-angiogenic factors. Nevertheless, they had a survival-enhancing effect on endothelial cells that might be useful in a variety of cell therapy or tissue-engineering approaches.

PMID: 21762016 [PubMed - as supplied by publisher]


Human adipose tissue derived mesenchymal stem cells are resistant to several chemotherapeutic agents.
Source:  NIH.gov
Monday, 29 November 1999 17:00

Human adipose tissue derived mesenchymal stem cells are resistant to several chemotherapeutic agents.

Cytotechnology. 2011 Oct;63(5):523-30

Authors: Liang W, Xia H, Li J, Zhao RC

Abstract
Human adipose derived mesenchymal stem cells (ADMSCs) are multipotential stem cells, originated from the vascular stromal compartment of fat tissues which can be used as an alternative cell source for many different cell therapies. However, their response to chemotherapeutic agants remains unknown. Here we assessed the acute direct effects of individual chemotherapeutic drug on ADMSCs. Using an in vitro culture system, the response of ADMSCs to the three chemotherapeutic agents cisplatin, comptothecin and vincristine was determined in comparison with that of testicular germ cell tumour (TGCT) cell line. The recovery of cell numbers following exposure to chemotherapeutic agents were also evaluated. Our results showed that human ADMSCs were resistant to chemo-therapeutic agents which are commonly used in clinic, the full recovery was seen respectively in ADMSCs after the drug treatment. Moreover, ADMSCs maintained their stem cell characteristics in vitro after the exposure to all chemotherapeutic agents.

PMID: 21761127 [PubMed - in process]


Stromal cell-derived factor-1 enhances distraction osteogenesis-mediated skeletal tissue regeneration through the recruitment of endothelial precursors.
Source:  NIH.gov
Monday, 29 November 1999 17:00

Stromal cell-derived factor-1 enhances distraction osteogenesis-mediated skeletal tissue regeneration through the recruitment of endothelial precursors.

Bone. 2011 Oct;49(4):693-700

Authors: Fujio M, Yamamoto A, Ando Y, Shohara R, Kinoshita K, Kaneko T, Hibi H, Ueda M

Abstract
Distraction osteogenesis (DO) is a unique therapy that induces skeletal tissue regeneration without stem/progenitor cell transplantation. Although the self-regeneration property of DO provides many clinical benefits, the long treatment period required is a major drawback. A high-speed DO mouse model (H-DO), in which the distraction was done two times faster than in control DO (C-DO) mice, failed to generate new bone callus in the DO gap. We found that this was caused by the unsuccessful recruitment of bone marrow endothelial cells (BM-ECs)/endothelial progenitor cells (EPCs) into the gap. We then tested the ability of a local application of stromal cell-derived factor-1 (SDF-1), a major chemo-attractant for BM-ECs/EPCs, to accelerate the bone regeneration in H-DO. Our data showed that, in H-DO, SDF-1 induced callus formation in the gap through the recruitment of BM-ECs/EPCs, the maturation of neo-blood vessels, and increased blood flow. These results indicate that the active recruitment of endogenous BM-ECs/EPCs may provide a substantial clinical benefit for shortening the treatment period of DO.

PMID: 21741502 [PubMed - in process]


Plasminogen regulates stromal cell-derived factor-1/CXCR4-mediated hematopoietic stem cell mobilization by activation of matrix metalloproteinase-9.
Source:  NIH.gov
Monday, 29 November 1999 17:00

Plasminogen regulates stromal cell-derived factor-1/CXCR4-mediated hematopoietic stem cell mobilization by activation of matrix metalloproteinase-9.

Arterioscler Thromb Vasc Biol. 2011 Sep;31(9):2035-43

Authors: Gong Y, Fan Y, Hoover-Plow J

Abstract
OBJECTIVE: Granulocyte colony-stimulating factor (G-CSF) is a widespread therapeutic agent for stimulation of hematopoietic progenitor and stem cell (HPSC) mobilization from bone marrow (BM). Plasminogen (Plg) has been shown to be critical for HPSC mobilization. Here, we investigated the role of Plg in G-CSF-induced HPSC mobilization and the underlying mechanism.
METHODS AND RESULTS: By using gene-targeted mice, our data show that Plg is required for G-CSF-induced HPSC egress to sinusoidal capillaries in BM and subsequent mobilization to peripheral circulation. G-CSF induced Plg-dependent activation of matrix metalloproteinase-9 (MMP-9) in BM, and MMP-9 neutralization or deficiency suppressed HPSC migration and mobilization. Reconstitution of MMP-9 activity by BM transplantation after lentiviral overexpression rescued HPSC mobilization in Plg-deficient mice, indicating that MMP-9 activation is required for Plg-mediated HPSC mobilization. Interestingly, after G-CSF simulation, Plg downregulated stromal cell-derived factor-1 in BM and spatiotemporally regulated the expression of C-X-C chemokine receptor type 4 (CXCR4) on mobilized HPSCs, and reconstitution of MMP-9 activity in Plg-deficient mice reversed CXCR4 expression on HPSCs in plasma and BM, suggesting that CXCR4 serves as a new downstream signal of Plg/MMP-9 in HPSC mobilization.
CONCLUSIONS: Our data elucidated a novel mechanism that Plg regulates MMP-9-dependent CXCR4 expression to facilitate HPSC mobilization in response to G-CSF.

PMID: 21719761 [PubMed - indexed for MEDLINE]


Regeneration of dental pulp by stem cells.
Source:  NIH.gov
Monday, 29 November 1999 17:00

Regeneration of dental pulp by stem cells.

Adv Dent Res. 2011 Jul;23(3):313-9

Authors: Nakashima M, Iohara K

Abstract
Angiogenesis/vasculogenesis and neurogenesis are essential for pulp regeneration. Two subfractions of side-population (SP) cells, CD31(-)/CD146(-) SP cells and CD105(+) cells with angiogenic and neurogenic potential, were isolated by flow cytometry from canine dental pulp. In an experimental model of mouse hindlimb ischemia, transplantation of these cell populations resulted in an increase in blood flow, including high-density capillary formation. In a model of rat cerebral ischemia, stem cell transplantations enhanced neuronal regeneration and recovery from motor disability. Autologous transplantation of the CD31(-)/CD146(-) SP cells into an in vivo model of amputated pulp resulted in complete regeneration of pulp tissue with vascular and neuronal processes within 14 days. The transplanted cells expressed pro-angiogenic factors, implying trophic action on endothelial cells. Autologous transplantation of CD31(-)/CD146(-) SP cells or CD105(+) cells with stromal-cell-derived factor-1 (SDF-1) into root canals after whole pulp removal of mature teeth resulted in complete regeneration of pulp replete with nerves and vasculature by day 14, followed by dentin formation along the dentinal wall by day 35. Therefore, the potential utility of fractionated SP cells and CD105(+) cells in angiogenesis and neurogenesis was demonstrated by treatment of limb and cerebral ischemia following pulpotomy and pulpectomy.

PMID: 21677085 [PubMed - indexed for MEDLINE]


Sonic hedgehog-induced functional recovery after myocardial infarction is enhanced by AMD3100-mediated progenitor-cell mobilization.
Source:  NIH.gov
Monday, 29 November 1999 17:00

Sonic hedgehog-induced functional recovery after myocardial infarction is enhanced by AMD3100-mediated progenitor-cell mobilization.

J Am Coll Cardiol. 2011 Jun 14;57(24):2444-52

Authors: Roncalli J, Renault MA, Tongers J, Misener S, Thorne T, Kamide C, Jujo K, Tanaka T, Ii M, Klyachko E, Losordo DW

Abstract
OBJECTIVES: This study was designed to compare the effectiveness of Sonic hedgehog (Shh) gene transfer, AMD3100-induced progenitor-cell mobilization, and Shh-AMD3100 combination therapy for treatment of surgically induced myocardial infarction (MI) in mice.
BACKGROUND: Shh gene transfer improves myocardial recovery by up-regulating angiogenic genes and enhancing the incorporation of bone marrow-derived progenitor cells (BMPCs) in infarcted myocardium. Here, we investigated whether the effectiveness of Shh gene therapy could be improved with AMD3100-induced progenitor-cell mobilization.
METHODS: Gene expression and cell function were evaluated in cells cultured with medium collected from fibroblasts transfected with plasmids encoding human Shh (phShh). MI was induced in wild-type mice, in matrix metalloproteinase (MMP)-9 knockout mice, and in mice transplanted with bone marrow that expressed green-fluorescent protein. Mice were treated with 100 μg of phShh (administered intramyocardially), 5 mg/kg of AMD3100 (administered subcutaneously), or both; cardiac function was evaluated echocardiographically, and fibrosis, capillary density, and BMPC incorporation were evaluated immunohistochemically.
RESULTS: phShh increased vascular endothelial growth factor and stromal cell-derived factor 1 expression in fibroblasts; the medium from phShh-transfected fibroblasts increased endothelial-cell migration and the migration, proliferation, and tube formation of BMPCs. Combination therapy enhanced cardiac functional recovery (i.e., left ventricular ejection fraction) in wild-type mice, but not in MMP-9 knockout mice, and was associated with less fibrosis, greater capillary density and smooth muscle-containing vessel density, and enhanced BMPC incorporation.
CONCLUSIONS: Combination therapy consisting of intramyocardial Shh gene transfer and AMD3100-induced progenitor-cell mobilization improves cardiac functional recovery after MI and is superior to either individual treatment for promoting therapeutic neovascularization.

PMID: 21658566 [PubMed - indexed for MEDLINE]


Stromal cell-derived factor-1 retention and cardioprotection for ischemic myocardium.
Source:  NIH.gov
Monday, 29 November 1999 17:00

Stromal cell-derived factor-1 retention and cardioprotection for ischemic myocardium.

Circ Heart Fail. 2011 Jul;4(4):509-18

Authors: Kanki S, Segers VF, Wu W, Kakkar R, Gannon J, Sys SU, Sandrasagra A, Lee RT

Abstract
BACKGROUND: Stromal cell-derived factor-1 (SDF-1) is a chemoattractant of stem/progenitor cells, and several studies have shown that SDF-1 may improve ventricular function after infarction. SDF-1 is cleaved by proteases including matrix metalloproteinase-2 (MMP-2) and CD26/dipeptidylpeptidase-4 (DPP-4), which are activated in injured tissues.
METHODS AND RESULTS: We investigated the biodistribution and functional roles of SDF-1 in experimental ischemia/reperfusion injury in rats. Radiolabeled SDF-1 given by intracoronary injection was selectively concentrated in ischemic myocardium. The enhanced uptake of SDF-1 in ischemic myocardium was not mediated by its receptor, CXCR4. Mass spectrometry and Western analyses showed that SDF-1 was cleaved by DPP-4 in plasma and myocardium, whereas a bioengineered MMP-2/DPP-4-resistant form of SDF-1, SSDF-1(S4V), was highly stable. A single dose of SSDF-1(S4V) exhibited greater potency for cardioprotection than wild-type SDF-1. SSDF-1(S4V) improved cardiac function in rats even after a 3-hour ischemic period.
CONCLUSIONS: These results show that a single dose of protease-resistant SSDF-1(S4V) after myocardial infarction leads to dramatic improvement in angiogenesis and ventricular function even 3 hours after the onset of ischemia, revealing a simple, clinically feasible approach to prevention of heart failure.

PMID: 21606214 [PubMed - indexed for MEDLINE]


A 3D in vitro bone organ model using human progenitor cells.
Source:  NIH.gov
Monday, 29 November 1999 17:00

A 3D in vitro bone organ model using human progenitor cells.

Eur Cell Mater. 2011;21:445-58; discussion 458

Authors: Papadimitropoulos A, Scherberich A, Güven S, Theilgaard N, Crooijmans HJ, Santini F, Scheffler K, Zallone A, Martin I

Abstract
Three-dimensional (3D) organotypic culture models based on human cells may reduce the use of complex and costly animal models, while gaining clinical relevance. This study aimed at developing a 3D osteoblastic-osteoclastic-endothelial cell co-culture system, as an in vitro model to mimic the process of bone turnover. Osteoprogenitor and endothelial lineage cells were isolated from the stromal vascular fraction (SVF) of human adipose tissue, whereas CD14+ osteoclast progenitors were derived from human peripheral blood. Cells were co-cultured within 3D porous ceramic scaffolds using a perfusion-based bioreactor device, in the presence of typical osteoclastogenic factors. After 3 weeks, the scaffolds contained cells with endothelial (2.0±0.3%), pre/osteoclastic (14.0±1.4%) and mesenchymal/osteoblastic (44.0±8.4%) phenotypes, along with tartrate-resistant acid phosphatase-positive (TRAP+) osteoclastic cells in contact with deposited bone-like matrix. Supernatant analysis demonstrated sustained matrix deposition (by C-terminus procollagen-I propeptides), resorption (by N-terminus collagen-I telopeptides and phosphate levels) and osteoclastic activity (by TRAP-5b) only when SVF and CD14+ cells were co-cultured. Scanning electron microscopy and magnetic resonance imaging confirmed the pattern of matrix deposition and resorption. The effectiveness of Vitamin D in replacing osteoclastogenic factors indicated a functional osteoblast-osteoclast coupling in the system. The formation of human-origin bone-like tissue, blood vessels and osteoclasts upon ectopic implantation validated the functionality of the developed cell types. The 3D co-culture system and the associated non-invasive analytical tools can be used as an advanced model to capture some aspects of the functional coupling of bone-like matrix deposition and resorption and could be exploited toward the engineering of multi-functional bone substitute implants.

PMID: 21604244 [PubMed - indexed for MEDLINE]


Growth of engineered human myocardium with mechanical loading and vascular coculture.
Source:  NIH.gov
Monday, 29 November 1999 17:00

Growth of engineered human myocardium with mechanical loading and vascular coculture.

Circ Res. 2011 Jun 24;109(1):47-59

Authors: Tulloch NL, Muskheli V, Razumova MV, Korte FS, Regnier M, Hauch KD, Pabon L, Reinecke H, Murry CE

Abstract
RATIONALE: The developing heart requires both mechanical load and vascularization to reach its proper size, yet the regulation of human heart growth by these processes is poorly understood.
OBJECTIVE: We seek to elucidate the responses of immature human myocardium to mechanical load and vascularization using tissue engineering approaches.
METHODS AND RESULTS: Using human embryonic stem cell and human induced pluripotent stem cell-derived cardiomyocytes in a 3-dimensional collagen matrix, we show that uniaxial mechanical stress conditioning promotes 2-fold increases in cardiomyocyte and matrix fiber alignment and enhances myofibrillogenesis and sarcomeric banding. Furthermore, cyclic stress conditioning markedly increases cardiomyocyte hypertrophy (2.2-fold) and proliferation rates (21%) versus unconditioned constructs. Addition of endothelial cells enhances cardiomyocyte proliferation under all stress conditions (14% to 19%), and addition of stromal supporting cells enhances formation of vessel-like structures by ≈10-fold. Furthermore, these optimized human cardiac tissue constructs generate Starling curves, increasing their active force in response to increased resting length. When transplanted onto hearts of athymic rats, the human myocardium survives and forms grafts closely apposed to host myocardium. The grafts contain human microvessels that are perfused by the host coronary circulation.
CONCLUSIONS: Our results indicate that both mechanical load and vascular cell coculture control cardiomyocyte proliferation, and that mechanical load further controls the hypertrophy and architecture of engineered human myocardium. Such constructs may be useful for studying human cardiac development as well as for regenerative therapy.

PMID: 21597009 [PubMed - indexed for MEDLINE]


Expansion of the human adipose-derived stromal vascular cell fraction yields a population of smooth muscle-like cells with markedly distinct phenotypic and functional properties relative to mesenchymal stem cells.
Source:  NIH.gov
Monday, 29 November 1999 17:00

Expansion of the human adipose-derived stromal vascular cell fraction yields a population of smooth muscle-like cells with markedly distinct phenotypic and functional properties relative to mesenchymal stem cells.

Tissue Eng Part C Methods. 2011 Aug;17(8):843-60

Authors: Basu J, Genheimer CW, Guthrie KI, Sangha N, Quinlan SF, Bruce AT, Reavis B, Halberstadt C, Ilagan RM, Ludlow JW

Abstract
Adipose tissue contains a heterogeneous cell population composed of endothelial cells, adipocytes, smooth muscle cells (SMC), and mesenchymal progenitors and stromal cells that meet the criteria put forth by the International Society for Cellular Therapy as defining mesenchymal stem cells (MSC). In this study, we expanded the stromal vascular fraction (SVF) of human adipose tissue and characterized the resulting adherent primary cell cultures by quantitative reverse transcription-polymerase chain reaction, antigen expression, protein fingerprinting, growth kinetics, in vitro tri-lineage differentiation bioactivity, and functional responses to small molecules modulating SMC-related developmental pathways and compared the results to those obtained with functionally validated MSC cultures. SVF-derived initial cultures (P0) were expanded in a defined medium that was not optimized for MSC growth conditions, neither were recombinant cytokines or growth factors added to the media to direct differentiation. The adherent cell cultures derived from SVF expansion under these conditions had markedly distinct phenotypic and biological properties relative to functionally validated MSC cultures. SVF-derived adherent cell cultures retained characteristics consistent with the SMC subpopulation within adipose tissue--phenotype, gene, and protein expression--that were independent of passage number and source of SVF (n=4 independent donors). SVF-derived cells presented significantly less robust in vitro tri-lineage differentiation bioactivity relative to validated MSC. Expanded SVF cells and MSC had opposite responses to the thromboxane A2 mimetic U46619, demonstrating an unambiguous functional distinction between the two cell types. Taken together, these data support the conclusions that SVF cells expanded under the conditions described in these studies are accurately described as adipose-derived SMC and represent a cellular subpopulation of adipose SVF that is separate and distinct from other classes of adipose-derived cells.

PMID: 21595545 [PubMed - in process]


Clinical trial of autologous differentiated adipocytes from stem cells derived from human adipose tissue.
Source:  NIH.gov
Monday, 29 November 1999 17:00

Clinical trial of autologous differentiated adipocytes from stem cells derived from human adipose tissue.

Dermatol Surg. 2011 Jun;37(6):750-9

Authors: Kim M, Kim I, Lee SK, Bang SI, Lim SY

Abstract
BACKGROUND: Adipose tissue-derived stem cells (ASCs) are considered to be a reliable cell source for the generation of adipose tissue because they can be differentiated into adipocytes. Previous data have indicated that adipogenic differentiation of ASCs before transplantation can enhance the regeneration of adipose tissue.
OBJECTIVE: To evaluate the efficacy and safety of the use of autologous differentiated adipocytes for the treatment of depressed scars.
METHODS: Autologous differentiated adipocytes were produced using well-established techniques, including the harvesting of stromal vascular fraction cells from lipoaspirates, expansion of ASCs, and differentiation into adipocytes. This was an open-label, dose-escalation study. Patients were given a subcutaneous injection of differentiated adipocytes and followed for 12 weeks.
RESULTS: Thirty-one patients were injected with differentiated adipocytes. When the differentiated adipocytes were injected subcutaneously into depressed scars, the average recovery in volume was 74.6% at 12 weeks. Of 17 patients who completed the follow-up after determination of dose, seven were willing to enroll for extended follow-up. Long-term follow-up revealed that the recovery in volume at 12 weeks was maintained well for at least 1 year. There were no significant adverse events.
CONCLUSIONS: The use of autologous differentiated adipocytes can be a safe and effective treatment for soft tissue defects, with relatively long-term maintenance of volume. The authors have indicated no significant interest with commercial supporters.

PMID: 21575095 [PubMed - indexed for MEDLINE]


The Rho kinase inhibitor fasudil augments the number of functional endothelial progenitor cells in ex vivo cultures.
Source:  NIH.gov
Monday, 29 November 1999 17:00

The Rho kinase inhibitor fasudil augments the number of functional endothelial progenitor cells in ex vivo cultures.

Int J Mol Med. 2011 Sep;28(3):357-63

Authors: O E, Ahn HY, Kim HK, You JC, Shin JC, Joe YA

Abstract
Rho kinase (ROCK) has been implicated in the regulation of vascular tone, endothelial dysfunction, inflammation and remodeling. Endothelial progenitor cells (EPC) have been proven to have the efficacy of therapeutic neovascularization in ischemia. However, the scarcity of EPCs limits cell therapy. Using an in vitro EPC culture assay, Y27632 was found to increase the number of adherent EPCs. In this study, we investigated the effect of fasudil, another ROCK inhibitor being used in the clinic, on EPC number and examined whether EPCs expanded by fasudil are functional in vitro and in vivo. In ex vivo cultures of EPCs, fasudil effectively increased the number of ac-LDL/UEA-1 positive cells as well as adherent cells, in contrast to H89, a less selective ROCK inhibitor. Fasudil also increased EPC numbers in culture up to 10 µM, in a dose-dependent manner. When EPCs expanded with fasudil were examined for the migratory activity toward stromal cell-derived factor-1 and vascular endothelial growth factor, these cells retained functional properties in migration, albeit with some decrease. Fasudil-cultured EPCs labeled with PKH26 showed an activity similar to non-treated EPCs for cellular adhesion into an endothelial cell (EC) monolayer and incorporation into capillary-like structures formed by ECs. Finally, when EPCs cultured with fasudil (106 cells/mouse) were injected into ischemic limbs, these cells showed a blood flow recovery at a level comparable to non-treated control EPCs and increased neovascularization. Therefore, these data suggest that the ROCK inhibitor fasudil can provide a beneficial effect in the treatment of ischemic diseases by increasing EPC numbers.

PMID: 21567077 [PubMed - indexed for MEDLINE]


New cell therapy using bone marrow-derived stem cells/endothelial progenitor cells to accelerate neovascularization in healing of experimental ulcerative colitis.
Source:  NIH.gov
Monday, 29 November 1999 17:00

New cell therapy using bone marrow-derived stem cells/endothelial progenitor cells to accelerate neovascularization in healing of experimental ulcerative colitis.

Curr Pharm Des. 2011;17(16):1643-51

Authors: Deng X, Szabo S, Chen L, Paunovic B, Khomenko T, Tolstanova G, Tarnawski AS, Jones MK, Sandor Z

Abstract
Inflammatory bowel disease (IBD): ulcerative colitis (UC) and Crohn disease (CD) are characterized by recurrent inflammation and ulceration of intestinal and/or colonic mucosa and an inappropriate and delayed healing. Current therapies with, e.g., anti-TNFα antibody (infliximab) and other anti-inflammatory drugs (e.g., mesalamine) do not induce sustained remission, complete healing or prevent recurrence of UC. Although the pathogenesis of UC is not fully understood, pathologic angiogenesis has been postulated as a critical pathogenic component in UC. Recent studies demonstrated that the poor healing, chronic inflammation in colon of UC could be the result of microvascular dysfunction and endothelial barrier defect, resulting in sustained tissue hypoperfusion and ischemia in the colon. Previously, regeneration of injured endothelium and neovascularization were believed to rely solely on the migration and proliferation of neighboring endothelial cells from existing blood vessels. However, accumulating evidence shows that additional mechanisms may exist, and may be mediated by the circulating pool of bone marrow-derived endothelial progenitor cells (BMD-EPC). Furthermore, stromal cell-derived factor-1 (SDF-1) and its receptor CXCR4 have been demonstrated to play an important role in the "homing" of BMD-EPC to injured sites and neovascularization in tissue repair. Recent studies by others and us showed reduced BMD-EPC levels in the circulation of IBD patients and rats with experimental UC. However, the potential therapeutic effect of BMD-EPC on neovascularization and colonic mucosal repair in UC has not been elucidated. In this review, we discussed the possibility that impaired contribution of BMD-EPC (i.e., decreased release of BMD-EPC from bone marrow to circulation and/or blocked/impaired homing of BMD-EPC to colonic lesions) may be a critical component of mechanisms in the incomplete/delayed healing of UC, and may offer a novel form of cell therapy for IBD.

PMID: 21548863 [PubMed - in process]


Myelosuppression of thrombocytes and monocytes is associated with a lack of synergy between chemotherapy and anti-VEGF treatment.
Source:  NIH.gov
Monday, 29 November 1999 17:00

Myelosuppression of thrombocytes and monocytes is associated with a lack of synergy between chemotherapy and anti-VEGF treatment.

Neoplasia. 2011 May;13(5):419-27

Authors: Starlinger P, Brugger P, Schauer D, Sommerfeldt S, Tamandl D, Kuehrer I, Schoppmann SF, Gnant M, Brostjan C

Abstract
PURPOSE: Chemotherapeutic agents that have shown improved patient outcome when combined with anti-vascular endothelial growth factor (VEGF) therapy were recently identified to induce the mobilization of proangiogenic Tie-2-expressing monocytes (TEMs) and endothelial progenitor cells (EPCs) by platelet release of stromal cell-derived factor 1α (SDF-1α). VEGF blockade was found to counteract cell mobilization. We aimed to determine why agents like gemcitabine do not elicit TEM and EPC recruitment and may therefore lack synergy with anti-VEGF therapy.
EXPERIMENTAL DESIGN: Locally advanced pancreatic cancer patients (n = 20) were monitored during 16 weeks of neoadjuvant therapy. Treatment was based on gemcitabine with or without the addition of bevacizumab. Blood levels of proangiogenic cell populations and angiogenesis factors were determined in 2-week intervals.
RESULTS: The lack of EPC mobilization during gemcitabine therapy was associated with severe thrombocytopenia and reduced SDF-1α blood concentrations. Furthermore, myelosuppression by gemcitabine correlated significantly with loss of TEMs. With respect to angiogenic factors stored and released by platelets, plasma levels of the angiogenesis inhibitor thrombospondin 1 (TSP-1) were selectively decreased and correlated significantly with thrombocytopenia in response to gemcitabine therapy.
CONCLUSIONS: A thorough literature screen identified thrombocytopenia as a common feature of chemotherapeutic agents that lack synergy with anti-VEGF treatment. Our results on gemcitabine therapy indicate that myelosuppression (in particular, with respect to thrombocytes and monocytes) interferes with the mobilization of proangiogenic cell types targeted by bevacizumab and may further counteract antiangiogenic therapy by substantially reducing the angiogenesis inhibitor TSP-1.

PMID: 21532882 [PubMed - indexed for MEDLINE]


Selecting the optimal cell for kidney regeneration: fetal, adult or reprogrammed stem cells.
Source:  NIH.gov
Monday, 29 November 1999 17:00

Selecting the optimal cell for kidney regeneration: fetal, adult or reprogrammed stem cells.

Organogenesis. 2011 Apr 1;7(2):123-34

Authors: Harari-Steinberg O, Pleniceanu O, Dekel B

Abstract
Chronic kidney disease (CKD) is a progressive loss in renal function over a period of months or years. End-stage renal disease (ESRD) or stage 5 CKD ensues when renal function deteriorates to under 15% of the normal range. ESRD requires either dialysis or, preferentially, a kidney organ allograft, which is severely limited due to organ shortage for transplantation. To combat this situation, one needs to either increase supply of organs or decrease their demand. Two strategies therefore exist: for those that have completely lost their kidney function (ESRD), we will need to supply new kidneys. Taking into account the kidneys' extremely complex structure, this may prove to be impossible in the near future. In contrast, for those patients that are in the slow progression route from CKD to ESRD but still have functional kidneys, we might be able to halt progression by introducing stem cell therapy to diseased kidneys to rejuvenate or regenerate individual cell types. Multiple cell compartments that fall into three categories are likely to be worthy targets for cell repair: vessels, stroma (interstitium) and nephron epithelia. Different stem/progenitor cells can be linked to regeneration of specific cell types; hematopoietic progenitors and hemangioblastic cell types have specific effects on the vascular niche (vasculogenesis and angiogenesis). Multipotent stromal cells (MSC), whether derived from the bone marrow or isolated from the kidney's non-tubular compartment, may, in turn, heal nephron epithelia via paracrine mechanisms. Nevertheless, as we now know that all of the above lack nephrogenic potential, we should continue our quest to derive genuine nephron (epithelial) progenitors from differentiated pluripotent stem cells, from fetal and adult kidneys and from directly reprogrammed somatic cells.

PMID: 21519195 [PubMed - in process]


Adipose injury-associated factors mitigate hypoxia in ischemic tissues through activation of adipose-derived stem/progenitor/stromal cells and induction of angiogenesis.
Source:  NIH.gov
Monday, 29 November 1999 17:00

Adipose injury-associated factors mitigate hypoxia in ischemic tissues through activation of adipose-derived stem/progenitor/stromal cells and induction of angiogenesis.

Am J Pathol. 2011 May;178(5):2322-32

Authors: Eto H, Suga H, Inoue K, Aoi N, Kato H, Araki J, Doi K, Higashino T, Yoshimura K

Abstract
Based on the analysis of exudates from injured adipose tissue, we prepared a mixture containing the injury-associated growth factors at the same proportion as the exudates, named adipose injury cocktail (AIC). We hypothesized that AIC induces a series of regenerating and angiogenic processes without actual wounding. The purpose of this study is to elucidate the therapeutic potentials of AIC. AIC preferentially activated adipose-derived stem/progenitor/stromal cells (ASCs) to proliferate, migrate, and form networks compared with vascular endothelial cells, whereas vascular endothelial growth factor did not induce mitogenesis or chemotaxis in human ASCs. Each component growth factor of AIC was differently responsible for the ASC activation. AIC-treated ASCs tended to differentiate into adipocytes or vessel-constituting cells rather than into other cell types. In ischemic adipose tissues of mice, induced by either a surgical intervention or diabetes, AIC administration enhanced proliferation, especially of CD31(-)/CD34(+) ASCs, and mitigated tissue hypoxia by increasing capillary density and reducing fibrogenesis. These results suggest that AIC may have therapeutic potentials for various ischemic/hypoxic conditions by inducing adipose remodeling and neovascularization through activation of ASCs and other cells. Treatment with AIC has many advantages over cell-based therapies regarding morbidity, cost, and physical risks and may be used as an alternative therapy for improving tissue oxygen.

PMID: 21514444 [PubMed - indexed for MEDLINE]


Effects of thalidomide on long-term bone marrow cultures from patients with myelodysplastic syndromes: induction of IL-10 expression in the stromal layers.
Source:  NIH.gov
Monday, 29 November 1999 17:00

Effects of thalidomide on long-term bone marrow cultures from patients with myelodysplastic syndromes: induction of IL-10 expression in the stromal layers.

Leuk Res. 2011 Aug;35(8):1102-7

Authors: Lazarini M, Traina F, Winnischofer SM, Costa FF, Queiroz ML, Saad ST

Abstract
The purpose of this study was to investigate the in vitro effects of thalidomide on long-term bone marrow cultures from patients with myelodysplastic syndrome. We demonstrated that thalidomide induced an increase in granulocyte-macrophage colony forming unit numbers and in IL-10 expression. Thalidomide also promoted a slight increase in IL-6, IL-1β and TNF-α expression in the stromal layers. The numbers of erythroid burst forming units, the apoptosis rate of hematopoietic cells, and VEGF and TNF-α expression levels in culture supernatants were not modulated. Our results indicate a participation of thalidomide upon the hematopoietic microenvironment of patients with myelodysplastic syndromes, especially in the up regulation of IL-10.

PMID: 21511336 [PubMed - indexed for MEDLINE]


Ceiling culture-derived proliferative adipocytes retain high adipogenic potential suitable for use as a vehicle for gene transduction therapy.
Source:  NIH.gov
Monday, 29 November 1999 17:00

Ceiling culture-derived proliferative adipocytes retain high adipogenic potential suitable for use as a vehicle for gene transduction therapy.

Am J Physiol Cell Physiol. 2011 Jul;301(1):C181-5

Authors: Asada S, Kuroda M, Aoyagi Y, Fukaya Y, Tanaka S, Konno S, Tanio M, Aso M, Satoh K, Okamoto Y, Nakayama T, Saito Y, Bujo H

Abstract
Adipose tissue is expected to provide a source of proliferative cells for regenerative medicine and cell-transplantation therapies using gene transfer manipulation. We have recently identified ceiling culture-derived proliferative adipocytes (ccdPAs) from the mature adipocyte fraction as cells suitable as a therapeutic gene vehicle because of their stable proliferative capacity. In this study, we examined the capability of adipogenic differentiation of the ccdPAs compared with stromal vascular fraction (SVF)-derived progenitor cells (adipose-derived stem cells, ASCs) with regard to their multipotential ability to be converted to another lineage and therefore their potential to be used for regenerative medicine research. After in vitro passaging, the surface antigen profile and the basal levels of adipogenic marker genes of the ccdPAs were not obviously different from those of the ASCs. However, the ccdPAs showed increased lipid-droplet accumulation accompanied with higher adipogenic marker gene expression after stimulation of differentiation compared with the ASCs. The higher adipogenic potential of the ccdPAs than the ASCs from the SVF was maintained for 42 days in culture. Furthermore, the difference in the adipogenic response was enhanced after partial stimulation without indomethacin. These results indicate that the ccdPAs retain a high adipogenic potential even after in vitro passaging, thus suggesting the commitment of ccdPAs to stable mature adipocytes after autotransplantation, indicating that they may have potential for use in regenerative and gene-manipulated medicine.

PMID: 21471463 [PubMed - indexed for MEDLINE]


Adult Neural Stem Cells: Response to Stroke Injury and Potential for Therapeutic Applications.
Source:  NIH.gov
Monday, 29 November 1999 17:00

Adult Neural Stem Cells: Response to Stroke Injury and Potential for Therapeutic Applications.

Curr Stem Cell Res Ther. 2011 Apr 5;

Authors: Barkho BZ, Zhao X

Abstract
The plasticity of neural stem/progenitor cells allows a variety of different responses to many environmental cues. In the past decade, significant research has gone into understanding the regulation of neural stem/progenitor cell properties, because of their promise for cell replacement therapies in adult neurological diseases. Both endogenous and grafted neural stem/progenitor cells are known to have the ability to migrate long distances to lesioned sites after brain injury and differentiate into new neurons. Several chemokines and growth factors, including stromal cell-derived factor-1 and vascular endothelial growth factor, have been shown to stimulate the proliferation, differentiation, and migration of neural stem/progenitor cells, and investigators have now begun to identify the critical downstream effectors and signaling mechanisms that regulate these processes. Both our own lab and others have shown that the extracellular matrix and matrix remodeling factors play a critical role in directing cell differentiation and migration of adult neural stem/progenitor cells within injured sites. Identification of these and other molecular pathways involved in stem cell homing into ischemic areas is vital for the development of new treatments. To ensure the best functional recovery, regenerative therapy may require the application of a combination approach that includes cell replacement, trophic support, and neural protection. Here we review the current state of our knowledge about endogenous adult and exogenous neural stem/progenitor cells as potential therapeutic agents for central nervous system injuries.

PMID: 21466483 [PubMed - as supplied by publisher]


Expansion of the Human Adipose-derived Stromal Vascular Cell Fraction Yields a Population of Smooth Muscle-like Cells with Markedly Distinct Phenotypic and Functional Properties Relative to Mesenchymal Stem Cells.
Source:  NIH.gov
Monday, 29 November 1999 17:00

Expansion of the Human Adipose-derived Stromal Vascular Cell Fraction Yields a Population of Smooth Muscle-like Cells with Markedly Distinct Phenotypic and Functional Properties Relative to Mesenchymal Stem Cells.

Tissue Eng Part C Methods. 2011 Apr 2;

Authors: Basu J, Genheimer C, Guthrie KI, Sangha N, Quinlan SF, Bruce AT, Reavis B, Halberstadt CR, Ilagan R, Ludlow JW

Abstract
Adipose tissue contains a heterogenous cell population composed of endothelial cells, adipocytes, smooth muscle cells (SMC), and mesenchymal progenitors and stromal cells that meet the criteria put forth by the International Society for Cellular Therapy (ISCT) as defining mesenchymal stem cells (MSC). In this study, we expanded the stromal vascular fraction (SVF) of human adipose tissue and characterized the resulting adherent primary cell cultures by quantitative reverse transcription polymerase chain reaction (qRT-PCR), antigen expression, protein fingerprinting, growth kinetics, in vitro tri-lineage differentiation bioactivity and functional responses to small molecules modulating smooth muscle cell related developmental pathways and compared the results to those obtained with functionally validated MSC cultures. SVF-derived initial cultures (P0) were expanded in defined media that was not optimized for MSC growth conditions, neither were recombinant cytokines or growth factors added to the media to direct differentiation. The adherent cell cultures derived from SVF expansion under these conditions had markedly distinct phenotypic and biological properties relative to functionally validated MSC cultures. SVF-derived adherent cell cultures retained characteristics consistent with the SMC subpopulation within adipose tissue - phenotype, gene and protein expression - that were independent of passage number and source of SVF (n=4 independent donors). SVF-derived cells presented significantly less robust in vitro tri-lineage differentiation bioactivity relative to validated MSC. Expanded SVF cells and MSC had opposite responses to the thromboxane A2 mimetic U46619, demonstrating an unambiguous functional distinction between the two cell types. Taken together, these data support the conclusions that SVF cells expanded under the conditions described in these studies are accurately described as adipose-derived smooth muscle-like cells (Ad-SMC) and represent a cellular subpopulation of adipose SVF that is separate and distinct from other classes of adipose-derived cells.

PMID: 21457102 [PubMed - as supplied by publisher]


[Mesenchymal stem cells: A therapeutic update].
Source:  NIH.gov
Monday, 29 November 1999 17:00

[Mesenchymal stem cells: A therapeutic update].

Med Sci (Paris). 2011 Mar;27(3):275-84

Authors: Jorgensen C, Deschaseaux F, Planat-Benard V, Gabison E

Abstract
Mesenchymal stem cells/multipotent marrow stromal cells (MSC) have the ability to participate in there construction of tissues both directly by providing repair cells (essentially those originating from mesoderm)and indirectly by modulating inflammatory and immune responses. This wide range of properties makes these cells very appealing to treat various pathological conditions. They have been first used in 1995 as supportive cells to facilitate hematopoietic stem cells engraftment, and then to minimize the deleterious consequences of graft versus host disease by their immunosuppressive function. Their robust osteogenic differentiation capacity has also been evaluated in numerous preclinical settings of healing/repair but more rarely in human clinical trials. During the past 10-15 years, the potential benefit of their paracrine actions has been tested in various situations such as to facilitate repair after cutaneous defects after burns or lower consequences of ischemic strokes. The purpose of this series of short texts is not to give an exhaustive panorama, but to discuss some well-identified indications in four different fields : auto-immune diseases,bone repair, vascular regeneration and eye lesions such as corneal and retinal defects.

PMID: 21447297 [PubMed - indexed for MEDLINE]


Concise review: Adipose-derived stromal vascular fraction cells and stem cells: let's not get lost in translation.
Source:  NIH.gov
Monday, 29 November 1999 17:00

Concise review: Adipose-derived stromal vascular fraction cells and stem cells: let's not get lost in translation.

Stem Cells. 2011 May;29(5):749-54

Authors: Gimble JM, Bunnell BA, Chiu ES, Guilak F

Abstract
Subcutaneous fat has emerged as an alternative tissue source for stromal/stem cells in regenerative medicine. Over the past decade, international research efforts have established a wealth of basic science and preclinical evidence regarding the differentiation potential and regenerative properties of both freshly processed, heterogeneous stromal vascular fraction cells and culture expanded, relatively homogeneous adipose-derived stromal/stem cells. The stage has been set for clinicians to translate adipose-derived cells from the bench to the bedside; however, this process will involve "development" steps that fall outside of traditional "hypothesis-driven, mechanism-based" paradigm. This concise review examines the next stages of the development process for therapeutic applications of adipose-derived cells and highlights the current state of the art regarding clinical trials. It is recommended that the experiments addressing these issues be reported comprehensively in the peer-review literature. This transparency will accelerate the standardization and reproducibility of adipose-derived cell therapies with respect to their efficacy and safety.

PMID: 21433220 [PubMed - indexed for MEDLINE]


Serum-deprived human multipotent mesenchymal stromal cells (MSCs) are highly angiogenic.
Source:  NIH.gov
Monday, 29 November 1999 17:00

Serum-deprived human multipotent mesenchymal stromal cells (MSCs) are highly angiogenic.

Stem Cell Res. 2011 May;6(3):215-25

Authors: Oskowitz A, McFerrin H, Gutschow M, Carter ML, Pochampally R

Abstract
Recent reports have indicated that mesenchymal stromal cells (MSCs) from bone marrow have a potential in vascular remodeling and angiogenesis. Here, we report a unique phenomenon that under serum-deprived conditions MSCs survive and replicate. Secretome analysis of MSCs grown under serum-deprived conditions (SD-MSCs) identified a significant upregulation of prosurvival and angiogenic factors including VEGF-A, ANGPTs, IGF-1, and HGF. An ex vivo rat aortic assay demonstrated longer neovascular sprouts generated from rat aortic rings cultured in SD-MSC-conditioned media compared to neovascular sprouts from aortas grown in MSC-conditioned media. With prolonged serum deprivation, a subpopulation of SD-MSCs began to exhibit an endothelial phenotype. This population expressed endothelial-specific proteins including VEGFR2, Tie2/TEK, PECAM/CD31, and eNOS and also demonstrated the ability to uptake acetylated LDL. SD-MSCs also exhibited enhanced microtubule formation in an in vitro angiogenesis assay. Modified chick chorioallantoic membrane (CAM) angiogenesis assays showed significantly higher angiogenic potential for SD-MSCs compared to MSCs. Analysis of CAMs grown with SD-MSCs identified human-specific CD31-positive cells in vascular structures. We conclude that under the stress of serum deprivation MSCs are highly angiogenic and a population of these cells has the potential to differentiate into endothelial-like cells.

PMID: 21421339 [PubMed - indexed for MEDLINE]


Management of mustard gas-induced limbal stem cell deficiency and keratitis.
Source:  NIH.gov
Monday, 29 November 1999 17:00

Management of mustard gas-induced limbal stem cell deficiency and keratitis.

Ophthalmology. 2011 Jul;118(7):1272-81

Authors: Javadi MA, Jafarinasab MR, Feizi S, Karimian F, Negahban K

Abstract
PURPOSE: To report the clinical findings and compare outcomes of different surgical techniques evolved over time in a large series of patients with delayed-onset mustard gas keratitis (MGK).
DESIGN: Retrospective, comparative, interventional case series.
PARTICIPANTS: Ninety Iranian male survivors (175 eyes) of Iraqi chemical warfare with chronic or delayed-onset MGK.
METHODS: The symptoms and clinical findings of patients are presented, and medical and surgical interventions to address dry eye, limbal ischemia and limbal stem cell deficiency (LSCD), and corneal involvements are explained. The results of limbal stem cell transplantation techniques (living-related conjunctival-limbal allograft [lrCLAL] versus keratolimbal allograft [KLAL]) as well as corneal transplantation techniques (penetrating keratoplasty [PK] versus lamellar keratoplasty [LK]) are compared in terms of clinical outcomes and graft survival rates.
MAIN OUTCOME MEASURES: Ocular findings and appropriate surgical approach for LSCD and corneal involvements.
RESULTS: A total of 175 eyes of 90 cases (all male) between 34 and 68 years of age were followed up for 101±30.3 months (range, 36-198 months). The most common ocular involvements were chronic blepharitis and dry eye. Conjunctival vascular abnormalities and limbal ischemia were observed in 27.4% and 29.7% of eyes, respectively. Limbal stem cell deficiency necessitating stem cell transplantation developed in 41.1% of eyes. The most common corneal sign was central and peripheral anterior stromal opacity (58.9%), followed by corneal stromal thinning (36.0%) and neovascularization (27.4%). Living-related conjunctival-limbal allograft was performed in 32 eyes, and KLAL was performed in 40 eyes. The rejection-free graft survival rate was 39.1% in the lrCLAL group and 80.7% in the KLAL group at month 40, with a mean length of 24.9 and 68.8 months, respectively (P = 0.02). Thirty eyes underwent PK and 51 underwent LK. Corneal graft failure was observed in 9 PK eyes and in 6 LK eyes. The rejection-free graft survival rate was 39.0% in the PK group and 90.3% in the LK group at month 28, with a mean length of 29.6 and 85.0 months, respectively (P<0.001).
CONCLUSIONS: Chemical warfare victims who initially have mild symptoms ultimately may experience significant ocular involvements requiring surgical intervention. Limbal and corneal abnormalities can be managed best by KLAL and LK, respectively.

PMID: 21397949 [PubMed - indexed for MEDLINE]


Mesenchymal stem cells restore lung function by recruiting resident and non-resident proteins.
Source:  NIH.gov
Monday, 29 November 1999 17:00

Mesenchymal stem cells restore lung function by recruiting resident and non-resident proteins.

Cell Transplant. 2011 Mar 7;

Authors: Jungebluth P, Luedde M, Ferrer E, Luedde T, Vucur M, Peinado VI, Go T, Schreiber C, Richthofen MV, Bader A, Haag J, Darsow KH, Bartel SJ, Lange HA, Furlani D, Steinhoff G, Macchiarini P

Abstract
Since human lungs are unlikely to repair or regenerate beyond the cellular level, cell therapy has not previously been considered for chronic irreversible obstructive lung diseases. To explore whether cell therapy can restore lung function, we administered allogenic intratracheal mesenchymal stem cell (MSC) in the trachea of rats with chronic thromboembolic pulmonary hypertension (CTEPH), a disease characterized by single or recurrent pulmonary thromboembolic obliteration and progressive pulmonary vascular remodeling. MSCs were retrieved only in high pressure-exposed lungs recruited via a homing stromal derived factor-1 alpha/CXCR4 pathway. After MSC administration, a marked and long-lasting improvement of all clinical parameters and a significant change of the proteome level were detected. Beside a variation of liver proteome, such as Caspase-3, NF- 〈B, Collagen1A1 and 〈-SMA, we also identified more than 300 resident and nonresident lung proteins, e.g. myosin light chain 3 (P16409) or mitochondrial ATP synthase subunit alpha (P15999). These results suggest that cell therapy restores lung function and the therapeutic effects of MSCs may be related to protein-based tissue reconstituting effects.

PMID: 21396162 [PubMed - as supplied by publisher]


Stromal vascular fraction from adipose tissue forms profound vascular network through the dynamic reassembly of blood endothelial cells.
Source:  NIH.gov
Monday, 29 November 1999 17:00

Stromal vascular fraction from adipose tissue forms profound vascular network through the dynamic reassembly of blood endothelial cells.

Arterioscler Thromb Vasc Biol. 2011 May;31(5):1141-50

Authors: Koh YJ, Koh BI, Kim H, Joo HJ, Jin HK, Jeon J, Choi C, Lee DH, Chung JH, Cho CH, Park WS, Ryu JK, Suh JK, Koh GY

Abstract
OBJECTIVE: Tremendous efforts have been made to establish effective therapeutic neovascularization using adipose tissue-derived stromal vascular fraction (SVF), but the efficiency is low, and underlying mechanisms and their interaction with the host in a new microenvironment are poorly understood.
METHODS AND RESULTS: Here we demonstrate that direct implantation of SVF derived from donor adipose tissue can create a profound vascular network through the disassembly and reassembly of blood endothelial cells at the site of implantation. This neovasculature successfully established connection with recipient blood vessels to form a functionally perfused circuit. Addition of vascular growth factors to the SVF implant improved the efficiency of functional neovasculature formation. In contrast, spheroid culture of SVF before implantation reduced the capacity of vasculature formation, possibly because of cellular alteration. Implanting SVF into the mouse ischemic hindlimb induced the robust formation of a local neovascular network and salvaged the limb. Moreover, the coimplantation of SVF prevented fat absorption in the subcutaneous adipose tissue graft model.
CONCLUSIONS: Freshly isolated SVF can effectively induce new vessel formation through the dynamic reassembly of blood endothelial cells and could be applied to achieve therapeutic neovascularization for relieving ischemia and preventing fat absorption in an autologous manner.

PMID: 21393582 [PubMed - indexed for MEDLINE]


Adipose-derived stromal/stem cells (ASC) in regenerative medicine: pharmaceutical applications.
Source:  NIH.gov
Monday, 29 November 1999 17:00

Adipose-derived stromal/stem cells (ASC) in regenerative medicine: pharmaceutical applications.

Curr Pharm Des. 2011;17(4):332-9

Authors: Gimble JM, Nuttall ME

Abstract
Information relating to the biology, culture expansion, and mechanisms relating to adipose-derived cells has advanced significantly in the past decade. Both the heterogeneous stromal vascular fraction (SVF) and more homogeneous adipose-derived stem cells (ASC) offer unique opportunities as novel cell-based therapeutics and as traditional pharmaceutical discovery tools. This review highlights the cytokine secretory functions of ASC and SVF cells as well as their potential use as immunomodulators and gene delivery vehicles. These functions make it feasible to exploit adipose-derived cells in the treatment of ischemic, musculoskeletal, and oncological disorders. With appropriate commercial development and in vivo validation, ASC and SVF cells will have a significant therapeutic impact in the future.

PMID: 21375497 [PubMed - indexed for MEDLINE]


Vascular gene transfer of SDF-1 promotes endothelial progenitor cell engraftment and enhances angiogenesis in ischemic muscle.
Source:  NIH.gov
Monday, 29 November 1999 17:00

Vascular gene transfer of SDF-1 promotes endothelial progenitor cell engraftment and enhances angiogenesis in ischemic muscle.

Mol Ther. 2011 May;19(5):895-902

Authors: Kuliszewski MA, Kobulnik J, Lindner JR, Stewart DJ, Leong-Poi H

Abstract
Gene therapy approaches to enhance endothelial progenitor cell (EPC) homing may augment cell engraftment to ischemic tissue and lead to a greater therapeutic response. Therefore, we assessed the effects of ultrasound-mediated (UM) transfection of the chemokine stromal cell-derived factor-1 (SDF-1) on homing and engraftment of intravenously administered EPCs and the subsequent angiogenic response in chronically ischemic skeletal muscle. Bone marrow-derived EPCs were isolated from donor Fisher 344 rats, cultured and labeled in preparation for injection into recipient animals via a jugular vein. Using a model of chronic hindlimb ischemia in rats, we demonstrated that UM destruction of intravenous carrier microbubbles loaded with SDF-1 plasmid DNA resulted in targeted transfection of the vascular endothelium within ischemic muscle and greater local engraftment of EPCs. The combination of SDF-1gene therapy and EPCs lead to the greatest increase in tissue perfusion and microvascular density within ischemic muscle, compared to no treatment or either monotherapy alone. Our results demonstrate that UM transfection of SDF-1 improves EPC targeting to chronically ischemic tissue, enhancing vascular engraftment and leading to a more robust neovascularization response.

PMID: 21364544 [PubMed - indexed for MEDLINE]


Regenerated silk fibroin scaffold and infrapatellar adipose stromal vascular fraction as feeder-layer: a new product for cartilage advanced therapy.
Source:  NIH.gov
Monday, 29 November 1999 17:00

Regenerated silk fibroin scaffold and infrapatellar adipose stromal vascular fraction as feeder-layer: a new product for cartilage advanced therapy.

Tissue Eng Part A. 2011 Jul;17(13-14):1725-33

Authors: Chlapanidas T, Faragò S, Mingotto F, Crovato F, Tosca MC, Antonioli B, Bucco M, Lucconi G, Scalise A, Vigo D, Faustini M, Marazzi M, Torre ML

Abstract
Articular cartilage has limited repair and regeneration potential, and the scarcity of treatment modalities has motivated attempts to engineer cartilage tissue constructs. The use of chondrocytes in cartilage tissue engineering has been restricted by the limited availability of these cells, their intrinsic tendency to lose their phenotype during the expansion, as well as the difficulties during the first cell adhesion to the scaffold. Aim of this work was to evaluate the intra-articular adipose stromal vascular fraction attachment on silk fibroin scaffold to promote chondrocytes adhesion and proliferation. Physicochemical characterization has demonstrated that three-dimensionally organized silk fibroin scaffold is an ideal biopolymer for cartilage tissue engineering; it allows cell attachment, scaffold colonization, and physically cell holding in the area that must be repaired; the use of adipose-derived stem cells is a promising strategy to promote adhesion and proliferation of chondrocytes to the scaffold as an autologous human feeder layer.

PMID: 21338265 [PubMed - indexed for MEDLINE]


Adult human adipose tissue contains several types of multipotent cells.
Source:  NIH.gov
Monday, 29 November 1999 17:00

Adult human adipose tissue contains several types of multipotent cells.

J Cardiovasc Transl Res. 2011 Apr;4(2):200-10

Authors: Tallone T, Realini C, Böhmler A, Kornfeld C, Vassalli G, Moccetti T, Bardelli S, Soldati G

Abstract
Multipotent mesenchymal stromal cells (MSCs) are a type of adult stem cells that can be easily isolated from various tissues and expanded in vitro. Many reports on their pluripotency and possible clinical applications have raised hopes and interest in MSCs. In an attempt to unify the terminology and the criteria to label a cell as MSC, in 2006 the International Society for Cellular Therapy (ISCT) proposed a standard set of rules to define the identity of these cells. However, MSCs are still extracted from different tissues, by diverse isolation protocols, are cultured and expanded in different media and conditions. All these variables may have profound effects on the selection of cell types and the composition of heterogeneous subpopulations, on the selective expansion of specific cell populations with totally different potentials and ergo, on the long-term fate of the cells upon in vitro culture. Therefore, specific molecular and cellular markers that identify MSCs subsets as well as standardization of expansion protocols for these cells are urgently needed. Here, we briefly discuss new useful markers and recent data supporting the rapidly emerging concept that many different types of progenitor cells are found in close association with blood vessels. This knowledge may promote the necessary technical improvements required to reduce variability and promote higher efficacy and safety when isolating and expanding these cells for therapeutic use. In the light of the discussed data, particularly the identification of new markers, and advances in the understanding of fundamental MSC biology, we also suggest a revision of the 2006 ISCT criteria.

PMID: 21327755 [PubMed - indexed for MEDLINE]


MDSC as a mechanism of tumor escape from sunitinib mediated anti-angiogenic therapy.
Source:  NIH.gov
Monday, 29 November 1999 17:00

MDSC as a mechanism of tumor escape from sunitinib mediated anti-angiogenic therapy.

Int Immunopharmacol. 2011 Jul;11(7):856-61

Authors: Finke J, Ko J, Rini B, Rayman P, Ireland J, Cohen P

Abstract
Sunitinib is a receptor tyrosine kinase inhibitor (TKI) that is front-line therapy for metastatic renal cell carcinoma (mRCC). Its antitumor activity is related to its ability to block tumor cell and tumor vasculature cell signaling via several TKI receptors (i.e. vascular endothelial growth factor receptors VEGFRs, platelet-derived growth factors (PDGFs), and stem cell factors). Sunitinib also targets myeloid derived suppressor cells (MDSCs) significantly reducing their accumulation in the peripheral blood and reversing T cell (IFNγ) suppression in both mRCC patients and in murine tumor models. This reduction in immune suppression provides a rationale for combining sunitinib with immunotherapy for the treatment of certain tumor types. Despite these encouraging findings, however, we have observed that sunitinib has variable impact at reducing MDSCs and restoring T cell function within the tumor microenvironment. Given the immunosuppressive and proangiogenic activities of MDSC, it seems plausible that their persistence may contribute to the resistance that develops in sunitinib-treated patients. While sunitinib reduced tumor infiltrating MDSCs in Renca and CT26-bearing mice, coinciding with strong to modest decreases in tumor size respectively, it was ineffective at reducing MDSCs (<35% reduction in Gr1+CD11b+) or tumor burden in 4T1-bearing mice. Persistence of intratumor MDSCs was paralleled by depressed intratumor T cell IFNγ response and increased GM-CSF expression. Additionally, in vitro and in vivo experiments showed that GM-CSF prolongs survival of MDSCs, thus protecting them from the effects of sunitinib via a pSTAT5-dependent pathway. Although preliminary, there is evidence of intratumor MDSC resistance in some mRCC patients following sunitinib treatment. Intratumor MDSC persistence and T cell IFNγ response post nephrectomy in patients receiving sunitinib in a neoadjuvant setting are being compared to RCC patients undergoing nephrectomy without prior sunitinib treatment. Tumors from untreated patients showed suppressed T cell IFNγ response along with substantial expression of MDSCs (5% of total digested cells). Thus far, tumors from 5/8 neoadjuvant patients showed persistence of intratumor MDSCs and low T cell IFNγ production post sunitinib treatment, findings that parallel results from untreated tumors. In the remaining 3 neoadjuvant patients, intratumor MDSCs were detected at low levels which coincided with a T cell IFNγ response similar to that observed with normal donor peripheral T cells. GM-CSF's role in promoting MDSC survival in patient tumors is supported by the observation that GM-CSF is produced in short-term RCC cultures at levels capable of protecting MDSCs from sunitinib-induced cell death. Additionally, persistence of MDSC also may be associated with increased expression of proangiogenic proteins, such as MMP9, MMP8, and IL-8 produced by tumor stromal cells or infiltrating MDSCs. Indeed our findings suggest that the most dominate MDSC subset in RCC patients is the neutrophilic population that produces proangiogenic proteins. We propose that the development of sunitinib resistance is partly mediated by the survival of MDSCs intratumorally, thereby providing sustained immune suppression and angiogenesis.

PMID: 21315783 [PubMed - indexed for MEDLINE]


Blood levels, apoptosis, and homing of the endothelial progenitor cells after skin burns and escharectomy.
Source:  NIH.gov
Monday, 29 November 1999 17:00

Blood levels, apoptosis, and homing of the endothelial progenitor cells after skin burns and escharectomy.

J Trauma. 2011 Feb;70(2):459-65

Authors: Foresta C, Schipilliti M, De Toni L, Magagna S, Lancerotto L, Azzena B, Vindigni V, Mazzoleni F

Abstract
BACKGROUND: Skin burns are an acute trauma involving an extensive vascular damage and an intense inflammatory response. Bone marrow-derived circulating endothelial progenitor cells (EPC) are known to migrate to sites of neovascularization in response to mediators (vascular endothelial growth factor and stromal cell-derived factor-1) released after trauma and ischemia, to contribute to wound healing, and to increase neovascularization of animal prefabricated flaps. Recent data showed an increase in EPC number in burned patients and a positive correlation between EPC number and total body surface area (TBSA) burnt, but data were limited to the first 5 days after thermal injury.
METHODS: By using flow cytometry, we studied EPC (CD34, CD133, CD45, and KDR cells) blood levels, apoptosis, and homing (stromal cell-derived factor-1 receptor expression and CXC chemokine receptor 4) in a 1-month follow-up postburn in 25 patients with ≥15% TBSA burnt, at least grade II burns and escharectomy performed at days 5 to 6, with respect to 31 controls.
RESULTS: EPC count at admission showed a positive linear correlation with TBSA burnt. The EPC blood levels of the patients were low (50.7 cells/mL±61.8 cells/mL) immediately after thermal injury, then increased with two peaks, at day 1 (188.3 cells/mL±223.2 cells/mL) and day 12 (253.1 cells/mL±430.7 cells/mL) with respect to controls (95.2 cells/mL±28.5 cells/mL, p<0.05), and then returned to normal levels in 1 month. EPC apoptotic rate and inflammatory parameters paralleled EPC blood count. No significant variations were found in CXC chemokine receptor 4 expression.
CONCLUSIONS: Thermal injury and escharectomy seem to induce an intense response in EPC production. In particular, escharectomy could improve physiologic wound repair by increasing EPC levels.

PMID: 21307748 [PubMed - indexed for MEDLINE]


Coronary vein infusion of multipotent stromal cells from bone marrow preserves cardiac function in swine ischemic cardiomyopathy via enhanced neovascularization.
Source:  NIH.gov
Monday, 29 November 1999 17:00

Coronary vein infusion of multipotent stromal cells from bone marrow preserves cardiac function in swine ischemic cardiomyopathy via enhanced neovascularization.

Lab Invest. 2011 Apr;91(4):553-64

Authors: Sato T, Iso Y, Uyama T, Kawachi K, Wakabayashi K, Omori Y, Soda T, Shoji M, Koba S, Yokoyama S, Fukuda N, Saito S, Katagiri T, Kobayashi Y, Takeyama Y, Umezawa A, Suzuki H

Abstract
Few reports have examined the effects of adult bone marrow multipotent stromal cells (MSCs) on large animals, and no useful method has been established for MSC implantation. In this study, we investigate the effects of MSC infusion from the coronary vein in a swine model of chronic myocardial infarction (MI). MI was induced in domestic swine by placing beads in the left coronary artery. Bone marrow cells were aspirated and then cultured to isolate the MSCs. At 4 weeks after MI, MSCs labeled with dye (n=8) or vehicle (n=5) were infused retrogradely from the anterior interventricular vein without any complications. Left ventriculography (LVG) was performed just before and at 4 weeks after cell infusion. The ejection fraction (EF) assessed by LVG significantly decreased from baseline up to a follow-up at 4 weeks in the control group (P<0.05), whereas the cardiac function was preserved in the MSC group. The difference in the EF between baseline and follow-up was significantly greater in the MSC group than in the control group (P<0.05). The MSC administration significantly promoted neovascularization in the border areas compared with the controls (P<0.0005), though it had no affect on cardiac fibrosis. A few MSCs expressed von Willebrand factor in a differentiation assay, but none of them expressed troponin T. In quantitative gene expression analysis, basic fibroblast growth factor and vascular endothelial growth factor (VEGF) levels were significantly higher in the MSC-treated hearts than in the controls (P<0.05, respectively). Immunohistochemical staining revealed VEGF production in the engrafted MSCs. In vitro experiment demonstrated that MSCs significantly stimulated endothelial capillary network formation compared with the VEGF protein (P<0.0001). MSC infusion via the coronary vein prevented the progression of cardiac dysfunction in chronic MI. This favorable effect appeared to derive not from cell differentiation, but from enhanced neovascularization by angiogenic factors secreted from the MSCs.

PMID: 21283079 [PubMed - indexed for MEDLINE]


Angiogenesis in ischemic tissue produced by spheroid grafting of human adipose-derived stromal cells.
Source:  NIH.gov
Monday, 29 November 1999 17:00

Angiogenesis in ischemic tissue produced by spheroid grafting of human adipose-derived stromal cells.

Biomaterials. 2011 Apr;32(11):2734-47

Authors: Bhang SH, Cho SW, La WG, Lee TJ, Yang HS, Sun AY, Baek SH, Rhie JW, Kim BS

Abstract
Stem cells offer significant therapeutic promise for the treatment of ischemic disease. However, stem cells transplanted into ischemic tissue exhibit limited therapeutic efficacy due to poor engraftment in vivo. Several strategies for improving the survival and engraftment of stem cells in ischemic tissue have been developed including transplantation in combination with growth factor delivery, genetic modification of stem cells, and the use of cell-transplantation scaffolds. Here, we demonstrate that human adipose-derived stromal cells (hADSCs) cultured and grafted as spheroids exhibit improved therapeutic efficacy for ischemia treatment. hADSCs were cultured in monolayer or spheroids. Spheroid cultures were more effective in preconditioning hADSCs to a hypoxic environment, upregulating hypoxia-adaptive signals (i.e., stromal cell-derived factor-1α and hypoxia-inducible factor-1α), inhibiting apoptosis, and enhancing secretion of both angiogenic and anti-apoptotic factors (i.e., hepatocyte growth factor, vascular endothelial growth factor, and fibroblast growth factor 2) compared to monolayer cultures. Moreover, cell harvesting following spheroid cultures avoided damage to extracellular matrices due to harsh proteolytic enzyme treatment, thereby preventing anoikis (apoptosis induced by a lack of cell-matrix interaction). Following intramuscular transplantation to ischemic hindlimbs of athymic mice, hADSC spheroids showed improved cell survival, angiogenic factor secretion, neovascularization, and limb survival as compared to hADSCs grafted as dissociated cells. Taken together, spheroid cultures precondition hADSCs to a hypoxic environment, and grafting hADSCs as spheroids to ischemic limbs improves therapeutic efficacy for ischemia treatment due to enhanced cell survival and paracrine effects. Spheroid-based cell delivery could be a simple and effective strategy for improving stem cell therapy for ischemic diseases, eliminating the need for growth factor delivery, biomaterial scaffolds or genetic modification.

PMID: 21262528 [PubMed - indexed for MEDLINE]


Intestinal myofibroblasts: targets for stem cell therapy.
Source:  NIH.gov
Monday, 29 November 1999 17:00

Intestinal myofibroblasts: targets for stem cell therapy.

Am J Physiol Gastrointest Liver Physiol. 2011 May;300(5):G684-96

Authors: Mifflin RC, Pinchuk IV, Saada JI, Powell DW

Abstract
The subepithelial intestinal myofibroblast is an important cell orchestrating many diverse functions in the intestine and is involved in growth and repair, tumorigenesis, inflammation, and fibrosis. The myofibroblast is but one of several α-smooth muscle actin-positive (α-SMA(+)) mesenchymal cells present within the intestinal lamina propria, including vascular pericytes, bone marrow-derived stem cells (mesenchymal stem cells or hematopoietic stem cells), muscularis mucosae, and the lymphatic pericytes (colon) and organized smooth muscle (small intestine) associated with the lymphatic lacteals. These other mesenchymal cells perform many of the functions previously attributed to subepithelial myofibroblasts. This review discusses the definition of a myofibroblast and reconsiders whether the α-SMA(+) subepithelial cells in the intestine are myofibroblasts or other types of mesenchymal cells, i.e., pericytes. Current information about specific, or not so specific, molecular markers of lamina propria mesenchymal cells is reviewed, as well as the origins of intestinal myofibroblasts and pericytes in the intestinal lamina propria and their replenishment after injury. Current concepts and research on stem cell therapy for intestinal inflammation are summarized. Information about the stem cell origin of intestinal stromal cells may inform future stem cell therapies to treat human inflammatory bowel disease (IBD).

PMID: 21252048 [PubMed - indexed for MEDLINE]


Rapid attachment of adipose stromal cells on resorbable polymeric scaffolds facilitates the one-step surgical procedure for cartilage and bone tissue engineering purposes.
Source:  NIH.gov
Monday, 29 November 1999 17:00

Rapid attachment of adipose stromal cells on resorbable polymeric scaffolds facilitates the one-step surgical procedure for cartilage and bone tissue engineering purposes.

J Orthop Res. 2011 Jun;29(6):853-60

Authors: Jurgens WJ, Kroeze RJ, Bank RA, Ritt MJ, Helder MN

Abstract
The stromal vascular fraction (SVF) of adipose tissue provides an abundant source of mesenchymal stem cells. For clinical application, it would be beneficial to establish treatments in which SVF is obtained, seeded onto a scaffold, and returned into the patient within a single surgical procedure. In this study, we evaluated the suitability of both a macroporous poly(L-lactide-co-caprolactone) and a porous collagen type I/III scaffold for this purpose. Surprisingly, cell attachment was rapid (∼10 min) and sequestered the majority of adipose stem cells, as deduced from colony-forming unit assays. Proliferation occurred in both polymeric scaffolds. Upon chondrogenic induction, up-regulation of chondrogenic genes, production of glycosaminoglycans, and accumulation of collagen type II was observed, indicating differentiation of scaffold-attached SVF cells along the chondrogenic lineage. Osteogenic differentiation was achieved in both scaffold types, as visualized by up-regulation of osteogenic genes, increase of alkaline phosphatase production over time, and accumulation of bone sialoprotein and osteonectin. In conclusion, this study identifies both poly(L-lactide-co-caprolactone) and collagen type I/III as promising scaffold materials for rapid attachment of adipose stem cell-like (stromal) cells, enhancing the development of one-step surgical concepts for cartilage and bone tissue engineering.

PMID: 21246614 [PubMed - indexed for MEDLINE]


SU5416 induces premature senescence in endothelial progenitor cells from patients with age-related macular degeneration.
Source:  NIH.gov
Monday, 29 November 1999 17:00

SU5416 induces premature senescence in endothelial progenitor cells from patients with age-related macular degeneration.

Mol Vis. 2011;17:85-98

Authors: Thill M, Berna MJ, Kunst F, Wege H, Strunnikova NV, Gordiyenko N, Grierson R, Richard G, Csaky KG

Abstract
PURPOSE: We recently demonstrated increased frequency and growth potential of late outgrowth endothelial progenitor cells (OECs) in patients with neovascular age-related macular degeneration (nvAMD). This study investigated the effects of short- and long-term in vitro inhibition of vascular endothelial growth factor (VEGF) Receptor-2 (VEGFR-2) signaling by SU5416 and other inhibitors of the VEGF signaling pathway in OECs.
METHODS: OECs, from the peripheral blood of patients with nvAMD, and human umbilical vein endothelial cells were grown in the presence of SU5416, other VEGFR-2 tyrosine kinase inhibitors (TKIs), and inhibitors of phosphatidylinositol 3'-Kinase (PI3K)/protein kinase B (Akt) and protein kinase C (PKC) in complete angiogenic medium. Apotosis was assessed after 48 h using the fluorescein isothiocyanate Annexin V method. Cell counts were performed for 10 days, and features of senescence were analyzed using senescence-associated β-galactosidase staining, the telomeric repeat amplification protocol for telomerase activity, Southern blot analysis for mean telomere length, flow cytometric analysis for cell-cycle arrest, and western blot for p53 and p21. Control OECs, cells treated for 7 days with inhibitors, as well as naturally senescent OECs were analyzed for expression of different endothelial antigens, including VEGFR-2 and the receptor for stromal cell-derived factor 1, chemokine receptor 4 (CXCR-4). Migration in vitro to VEGF and stromal cell-derived factor 1 of OECs was assessed.
RESULTS: SU5416, other VEGFR-2 TKIs, and inhibitors of PI3K, Akt, and PKC induced apoptosis, inhibited long-term proliferation, reduced telomerase activity, and induced premature senescence and cell-cycle arrest in OECs as well as in human umbilical vein endothelial cells. Naturally senescent cells and cells rendered senescent by VEGFR-2 TKIs had reduced VEGFR-2 and CXCR-4 expression and demonstrated reduced migratory ability to VEGF.
CONCLUSIONS: This study demonstrates apoptosis upon short-term inhibition and inhibition of long-term survival of OECs from patients with nvAMD by SU5416, presumably via PI3K/Akt and/or PKC-mediated reduction in telomerase activity and subsequent induction of premature senescence, which is accompanied by impaired endothelial activity. Therefore, induction of premature senescence in endothelial cells may represent a potential therapeutic target in nvAMD.

PMID: 21245959 [PubMed - indexed for MEDLINE]


Using living cells to transport therapeutic genes for cancer treatment.
Source:  NIH.gov
Monday, 29 November 1999 17:00

Using living cells to transport therapeutic genes for cancer treatment.

Clin Transl Oncol. 2011 Jan;13(1):10-7

Authors: Latorre-Romero C, Marin-Yaseli MR, Belmar-Lopez C, del Moral R, Marijuan PC, Quintanilla M, Martin-Duque P

Abstract
One of the key problems in cancer gene therapy is the inefficient delivery of therapeutic transgenes to tumour sites, after the systemic injection of the viral vector. Hence, new vector discovery is extremely important for the improvement of gene therapy results. Previously, mammalian cells were proposed as new vector systems; however with recent advances in stem cell research this modality makes them more suitable candidates. Tumours are composed of both malignant and benign cells. As "benign" cell types are able to form blood vessels, and stroma, it has been hypothesised that exogenously administrated cells of a different kind would preferentially engraft at the stromal tumour site and could deliver cancer gene therapy vectors to tumours.

PMID: 21239350 [PubMed - indexed for MEDLINE]


Angiogenesis after transplantation of auto- and allogenic cells.
Source:  NIH.gov
Monday, 29 November 1999 17:00

Angiogenesis after transplantation of auto- and allogenic cells.

Bull Exp Biol Med. 2010 Oct;149(4):457-61

Authors: Fatkhudinov TKh, Bol'shakova GB, Komissarova SV, Arutyunyan IV, Rzhaninova AA, Goldstein DV

Abstract
Neoangiogenesis after transplantation of auto- and allogenic mononuclears and multipotent stromal cells from the bone marrow was studied on the model of inflammatory angiogenesis. Transplanted auto- and allogenic cells stimulate the formation of new blood vessels in the granulation tissue, this manifesting in an increase in the quantity and volume density of blood vessels. The most pronounced angiogenesis was observed after transplantation of allogenic mononuclears and multipotent stromal cells. It was associated with intense inflammatory infiltration, with less numerous and mature collagen fibers in the granulation tissue. Injection of allogenic cells led to stimulation and chronization of inflammation, infiltration with inflammatory and poorly differentiated cells, and more pronounced and lasting angiogenesis. However, neither auto-, nor allogenic transplanted labeled cells were detected in the walls of new blood vessels. Hence, it seems that bone marrow mononuclears and multipotent stromal cells stimulated angiogenesis mainly at the expense of production of angiogenic factors, and after transplantation of allogenic cells also by stimulating the inflammation.

PMID: 21234443 [PubMed - indexed for MEDLINE]


Comparison of mesenchymal stem cells from adipose tissue and bone marrow for ischemic stroke therapy.
Source:  NIH.gov
Monday, 29 November 1999 17:00

Comparison of mesenchymal stem cells from adipose tissue and bone marrow for ischemic stroke therapy.

Cytotherapy. 2011 Jul;13(6):675-85

Authors: Ikegame Y, Yamashita K, Hayashi S, Mizuno H, Tawada M, You F, Yamada K, Tanaka Y, Egashira Y, Nakashima S, Yoshimura S, Iwama T

Abstract
BACKGROUND AIMS: Transplantation of mesenchymal stromal cells (MSC) derived from bone marrow (BM) or adipose tissue is expected to become a cell therapy for stroke. The present study compared the therapeutic potential of adipose-derived stem cells (ASC) with that of BM-derived stem cells (BMSC) in a murine stroke model.
METHODS: ASC and BMSC were isolated from age-matched C57BL/6J mice. These MSC were analyzed for growth kinetics and their capacity to secrete trophic factors and differentiate toward neural and vascular cell lineages in vitro. For in vivo study, ASC or BMSC were administrated intravenously into recipient mice (1 × 10(5) cells/mouse) soon after reperfusion following a 90-min middle cerebral artery occlusion. Neurologic deficits, the degree of infarction, expression of factors in the brain, and the fate of the injected cells were observed.
RESULTS: ASC showed higher proliferative activity with greater production of vascular endothelial cell growth factor (VEGF) and hepatocyte growth factor (HGF) than BMSC. Furthermore, in vitro conditions allowed ASC to differentiate into neural, glial and vascular endothelial cells. ASC administration showed remarkable attenuation of ischemic damage, although the ASC were not yet fully incorporated into the infarct area. Nonetheless, the expression of HGF and angiopoietin-1 in ischemic brain tissue was significantly increased in ASC-treated mice compared with the BMSC group.
CONCLUSIONS: Compared with BMSC, ASC have great advantages for cell preparation because of easier and safer access to adipose tissue. Taken together, our findings suggest that ASC would be a more preferable source for cell therapy for brain ischemia than BMSC.

PMID: 21231804 [PubMed - indexed for MEDLINE]


Comparison of mesenchymal stromal cells from young healthy donors and patients with severe chronic coronary artery disease.
Source:  NIH.gov
Monday, 29 November 1999 17:00

Comparison of mesenchymal stromal cells from young healthy donors and patients with severe chronic coronary artery disease.

Scand J Clin Lab Invest. 2011 May;71(3):193-202

Authors: Friis T, Haack-Sørensen M, Hansen SK, Hansen L, Bindslev L, Kastrup J

Abstract
BACKGROUND: It has been questioned whether bone marrow-derived mesenchymal stromal cells (MSCs) from patients with ischemic heart disease are suitable for use in regenerative stem cell therapy. We compared MSCs from patients with chronic coronary artery disease (CAD) and MSCs from young healthy donors with respect to phenotype, proliferation and endothelial differentiation capacity.
METHODS: MSCs from 16 young healthy donors and 15 elderly CAD patients were isolated, expanded by ex-vivo cultivation for two cell passages and characterized by flow cytometry, real time PCR and angiogenesis assay.
RESULTS: MSCs from healthy donors and CAD patients expressed the same surface markers and had similar proliferation capacity. In both groups VEGF-stimulation significantly increased the expression of the endothelial genes thrombospondin 1, Tie-2 and von Willebrand Factor and induced the capacity to form ring structures on extracellular matrix.
DISCUSSION: MSCs from young healthy donors and CAD patients proliferate equally well, express the same surface markers and increase in endothelial gene expression and ring structure formation capacity in the angiogenesis assay upon VEGF-stimulation. MSCs from CAD patients do not seem to be inferior to MSCs from young healthy donors thus indicating that autologous MSCs may be suitable for cell therapy in CAD patients.

PMID: 21222501 [PubMed - indexed for MEDLINE]


Adult stem cel diferentiation and trafficking and their implications in disease.
Source:  NIH.gov
Monday, 29 November 1999 17:00

Adult stem cel diferentiation and trafficking and their implications in disease.

Adv Exp Med Biol. 2010;695:169-83

Authors: Zhuge Y, Liu ZJ, Velazquez OC

Abstract
Stem cells are unspecialized precursor cells that mainly reside in the bone marrow and have important roles in the establishment of embryonic tissue. They also have critical functions during adulthood, where they replenish short-lived mature effector cells and regeneration of injured tissue. They have three main characteristics: self-renewal, differentiation and homeostatic control. In order to maintain a pool of stem cells that support the production of blood cells, stromal elements and connective tissue, stem cells must be able to constantly replenish their own number. They must also possess the ability to differentiate and give rise to a heterogeneous group of functional cells. Finally, stem cells must possess the ability to modulate and balance differentiation and self-renewal according to environmental stimuli and whole-organ needs to prevent the production of excessive number of effector cells.(1) In addition to formation of these cells, regulated movement of stem cells is critical for organogenesis, homeostasis and repair in adulthood. Stem cells require specific inputs from particular environments in order to perform their various functions. Some similar trafficking mechanisms are shared by leukocytes, adult and fetal stem cells, as well as cancer stem cells.(1,2) Achieving proper trafficking of stem cells will allow increased efficiency of targeted cell therapy and drug delivery.(2) In addition, understanding similarities and differences in homing and migration of malignant cancer stem cells will also clarify molecular events of tumor progression and metastasis.(2) This chapter focuses on the differentiation, trafficking and homing of the major types of adult bone marrow stem cells: hematopoietic stem cells (HSCs), mesenchymal stem cells (MSCs) and endothelial progenitor cells (EPCs) and the term"stem cell" will refer to "adult stem cells" unless otherwise specified.

PMID: 21222206 [PubMed - in process]


Human epicardium-derived cells fuse with high efficiency with skeletal myotubes and differentiate toward the skeletal muscle phenotype: a comparison study with stromal and endothelial cells.
Source:  NIH.gov
Monday, 29 November 1999 17:00

Human epicardium-derived cells fuse with high efficiency with skeletal myotubes and differentiate toward the skeletal muscle phenotype: a comparison study with stromal and endothelial cells.

Mol Biol Cell. 2011 Mar 1;22(5):581-92

Authors: Gentile A, Toietta G, Pazzano V, Tsiopoulos VD, Giglio AF, Crea F, Pompilio G, Capogrossi MC, Di Rocco G

Abstract
Recent studies have underscored a role for the epicardium as a source of multipotent cells. Here, we investigate the myogenic potential of adult human epicardium-derived cells (EPDCs) and analyze their ability to undergo skeletal myogenesis when cultured with differentiating primary myoblasts. Results are compared to those obtained with mesenchymal stromal cells (MSCs) and with endothelial cells, another mesodermal derivative. We demonstrate that EPDCs spontaneously fuse with pre-existing myotubes with an efficiency that is significantly higher than that of other cells. Although at a low frequency, endothelial cells may also contribute to myotube formation. In all cases analyzed, after entering the myotube, nonmuscle nuclei are reprogrammed to express muscle-specific genes. The fusion competence of nonmyogenic cells in vitro parallels their ability to reconstitute dystrophin expression in mdx mice. We additionally show that vascular cell adhesion molecule 1 (VCAM1) expression levels of nonmuscle cells are modulated by soluble factors secreted by skeletal myoblasts and that VCAM1 function is required for fusion to occur. Finally, treatment with interleukin (IL)-4 or IL-13, two cytokines released by differentiating myotubes, increases VCAM1 expression and enhances the rate of fusion of EPDCs and MSCs, but not that of endothelial cells.

PMID: 21209317 [PubMed - indexed for MEDLINE]


COMP-Ang1 stimulates HIF-1α-mediated SDF-1 overexpression and recovers ischemic injury through BM-derived progenitor cell recruitment.
Source:  NIH.gov
Monday, 29 November 1999 17:00

COMP-Ang1 stimulates HIF-1α-mediated SDF-1 overexpression and recovers ischemic injury through BM-derived progenitor cell recruitment.

Blood. 2011 Apr 21;117(16):4376-86

Authors: Youn SW, Lee SW, Lee J, Jeong HK, Suh JW, Yoon CH, Kang HJ, Kim HZ, Koh GY, Oh BH, Park YB, Kim HS

Abstract
Recruitment and adhesion of bone marrow (BM)-derived circulating progenitor cells to ischemic tissue are important for vasculogenesis and tissue repair. Recently, we found cartilage oligomeric matrix protein (COMP)-Ang1 is a useful cell-priming agent to improve the therapeutic efficacy of progenitor cells. However, the effect and the underlying mechanisms of COMP-Ang1 on recruitment of BM-derived progenitor cells (BMPCs) to foci of vascular injury have not been well defined. Here, we found that COMP-Ang1 is a critical stimulator of stromal cell-derived factor 1 (SDF-1), the principal regulator of BM-cell trafficking. Furthermore, SDF-1 stimulation by COMP-Ang1 was blocked by small-interfering RNA (siRNA) against hypoxia-inducible factor-1α (HIF-1α). COMP-Ang1 increased the synthesis of HIF-1α by activating mammalian target of rapamycin (mTOR) in hypoxic endothelium. The intermediate mechanism transmitting the COMP-Ang1 signal to the downstream mTOR/HIF-1α/SDF-1 pathway was the enhanced binding of the Tie2 receptor with integrin-linked kinase (ILK), an upstream activator of mTOR. In the mouse ischemic model, local injection of COMP-Ang1 stimulated the incorporation of BMPCs into ischemic limb, thereby enhancing neovasculogenesis and limb salvage. Collectively, our findings identify the COMP-Ang1/HIF-1α/SDF-1 pathway as a novel inducer of BMPC recruitment and neovasculogenesis in ischemic disease.

PMID: 21200018 [PubMed - indexed for MEDLINE]


Therapeutic effects of human STRO-3-selected mesenchymal precursor cells and their soluble factors in experimental myocardial ischemia.
Source:  NIH.gov
Monday, 29 November 1999 17:00

Therapeutic effects of human STRO-3-selected mesenchymal precursor cells and their soluble factors in experimental myocardial ischemia.

J Cell Mol Med. 2011 Oct;15(10):2117-29

Authors: See F, Seki T, Psaltis PJ, Sondermeijer HP, Gronthos S, Zannettino AC, Govaert KM, Schuster MD, Kurlansky PA, Kelly DJ, Krum H, Itescu S

Abstract
Stromal precursor antigen (STRO)-3 has previously been shown to identify a subset of adult human bone marrow (BM)-derived mesenchymal lineage precursors, which may have cardioprotective potential. We sought to characterize STRO-3-immunoselected and culture-expanded mesenchymal precursor cells (MPCs) with respect to their biology and therapeutic potential in myocardial ischemia. Immunoselection of STRO-3(+) MPCs enriched for fibroblastic colony forming units from unfractionated BM mononuclear cells (MNCs). Compared to mesenchymal stem cells conventionally isolated by plastic adherence, MPCs demonstrated increased proliferative capacity during culture expansion, expressed higher levels of early 'stem cell' markers and various pro-angiogenic and cardioprotective cytokines, and exhibited greater trilineage developmental efficiency. Intramyocardial injection of MPCs into a rat model of myocardial infarction (MI) promoted left ventricular recovery and inhibited left ventricular dilatation. These beneficial effects were associated with cardioprotective and pro-angiogenic effects at the tissue level, despite poor engraftment of cells. Treatment of MI rats with MPC-conditioned medium (CM) preserved left ventricular function and dimensions, reduced myocyte apoptosis and fibrosis, and augmented neovascularization, involving both resident vascular cells and circulating endothelial progenitor cells (EPCs). Profiling of CM revealed various cardioprotective and pro-angiogenic factors, which had biological activity in cultures of myocytes, tissue-resident vascular cells and EPCs. Prospective immunoselection of STRO-3(+) MPCs from BM MNCs conferred advantage in maintaining a population of immature MPCs during ex vivo expansion. Transplantation of culture-expanded MPCs into the post-MI heart resulted in therapeutic benefit, attributable at least in part to paracrine mechanisms of action. Thus, MPCs represent a promising therapy for myocardial ischemia.

PMID: 21155976 [PubMed - in process]


Bone marrow mesenchymal stromal cells with support of bispecific antibody and ultrasound-mediated microbubbles prevent myocardial fibrosis via the signal transducer and activators of transcription signaling pathway.
Source:  NIH.gov
Monday, 29 November 1999 17:00

Bone marrow mesenchymal stromal cells with support of bispecific antibody and ultrasound-mediated microbubbles prevent myocardial fibrosis via the signal transducer and activators of transcription signaling pathway.

Cytotherapy. 2011 Apr;13(4):431-40

Authors: Deng W, Chen QW, Li XS, Liu H, Niu SQ, Zhou Y, Li GQ, Ke DZ, Mo XG

Abstract
BACKGROUND AIMS: This study was initiated to investigate the efficacy of myocardial fibrosis intervention via signal transducer and activators of transcription (STAT) signaling using bone marrow (BM) mesenchymal stromal cells (MSC) with the aid of bispecific antibody (BiAb) and ultrasound-mediated microbubbles (MB).
METHODS: BiAb (anti-CD29 × anti-myosin light chain antibody; AMLCA) was prepared and combined with isolated MSC from male mice and transfused into female mice with isoproterenol-induced myocardial fibrosis via the tail vein, followed by MB (MSC + BiAb + MB). This study included seven groups: MSC + BiAb + MB; MSC; BiAb; MB; MSC + BiAb; untreated; and control. Five weeks after treatment, expression levels of the sex-determining region of Y-chromosome (SRY), matrix metalloproteinases (MMP)-9, tissue inhibitor of metalloproteinase (TIMP)-1 and vascular endothelial growth factor (VEGF) in myocardium were detected by fluorescent quantitative real-time polymerase chain reaction (qRT-PCR). Collagen distribution was observed using Sirius Red staining. The protein expression of signal transducer and activators of transcription (STAT)1 and STAT3 was detected by Western blot.
RESULTS: The highest homing number of MSC was in the MSC + BiAb + MB group, second highest in the MSC + BiAb group, and lowest in MSC alone. Compared with the untreated group, MSC + BiAb + MB, MSC + BiAb and MSC groups had decreased levels of MMP-9, TIMP-1, STAT1 and collagen deposition, and increased levels of STAT3. Upregulated STAT3 and downregulated TIMP-1 were significantly different in MSC + BiAb + MB compared with MSC alone or MSC + BiAb.
CONCLUSIONS: The homing rate and repairing efficacy of MSC improved with treatment utilizing a combination of BiAb and MB. MSC can improve MMP-TIMP expression in injured myocardium and interfere with myocardial fibrosis after homing, a mechanism that may be related to the STAT-mediated signaling pathway.

PMID: 21174489 [PubMed - indexed for MEDLINE]


Developing vasculature and stroma in engineered human myocardium.
Source:  NIH.gov
Monday, 29 November 1999 17:00

Developing vasculature and stroma in engineered human myocardium.

Tissue Eng Part A. 2011 May;17(9-10):1219-28

Authors: Kreutziger KL, Muskheli V, Johnson P, Braun K, Wight TN, Murry CE

Abstract
We recently developed a scaffold-free patch of human myocardium with human embryonic stem cell-derived cardiomyocytes and showed that stromal and endothelial cells form vascular networks in vitro and improve cardiomyocyte engraftment. Here, we hypothesize that stromal cells regulate the angiogenic phenotype by modulating the extracellular matrix (ECM). Human marrow stromal cells (hMSCs) support the greatest degree of endothelial cell organization, at 1.3- to 2.4-fold higher than other stromal cells tested. Stromal cells produce abundant ECM components in patches, including fibrillar collagen, hyaluronan, and versican. We identified two clonal hMSC lines that supported endothelial networks poorly and robustly. Interestingly, the pro-angiogenic hMSCs express high levels of versican, a chondroitin sulfate proteglycan that modulates angiogenesis and wound healing, whereas poorly angiogenic hMSCs produce little versican. When transplanted onto uninjured athymic rat hearts, patches with proangiogenic hMSCs develop ~ 50-fold more human vessels and form anastomoses with the host circulation, resulting in chimeric vessels containing erythrocytes. Thus, stromal cells play a key role in supporting vascularization of engineered human myocardium. Different stromal cell types vary widely in their proangiogenic ability, likely due in part to differences in ECM synthesis. Comparison of these cells defines an in vitro predictive platform for studying vascular development.

PMID: 21187004 [PubMed - indexed for MEDLINE]


Repeated and targeted transfer of angiogenic plasmids into the infarcted rat heart via ultrasound targeted microbubble destruction enhances cardiac repair.
Source:  NIH.gov
Monday, 29 November 1999 17:00

Repeated and targeted transfer of angiogenic plasmids into the infarcted rat heart via ultrasound targeted microbubble destruction enhances cardiac repair.

Eur Heart J. 2011 Aug;32(16):2075-84

Authors: Fujii H, Li SH, Wu J, Miyagi Y, Yau TM, Rakowski H, Egashira K, Guo J, Weisel RD, Li RK

Abstract
AIMS: Ultrasound-targeted microbubble destruction (UTMD) uses ultrasound energy to selectively deliver genes into the myocardium using plasmids conjugated to microbubbles. We hypothesized that repeated delivery of stem cell-mobilizing genes could boost the ability of this therapy to enhance cardiac repair and ventricular function after a myocardial infarction.
METHODS AND RESULTS: Beginning 7 days after coronary artery ligation, stem cell factor (SCF) and stromal cell-derived factor (SDF)-1α genes were administered to adult rats using 1, 3, or 6 UTMD treatments (repeat 1, 3, and 6 groups) at 2-day intervals (control=6 treatments with empty plasmid). Cardiac function (echocardiography) and myocardial perfusion (myocardial contrast echocardiography) were assessed on Days -7, 0, and 24 relative to the first treatment. Histological and biochemical assessments were performed on Day 24. Multiple UTMD treatments were associated with an increased presence of myocardial SCF and SDF-1α proteins and their receptors (vs. control and Repeat 1). All UTMD recipients exhibited increased vascular densities and smaller infarct regions (vs. control), with the highest ventricular densities in response to multiple treatments. Myocardial perfusion and ventricular function at Day 24 also improved progressively (vs. control) with the number of UTMD treatments.
CONCLUSIONS: Targeted ultrasound delivery of SCF and SDF-1α genes to the infarcted myocardium recruited progenitor cells and increased vascular density. Multiple UTMD treatments enhanced tissue repair, perfusion, and cardiac function. Repeated UTMD therapy may be applied to tailor the number of interventions required to optimize cardiac regeneration after an infarction.

PMID: 21196445 [PubMed - in process]


Efficient human fetal liver cell isolation protocol based on vascular perfusion for liver cell based therapy and a case report on cell transplantation.
Source:  NIH.gov
Monday, 29 November 1999 17:00

Efficient human fetal liver cell isolation protocol based on vascular perfusion for liver cell based therapy and a case report on cell transplantation.

Liver Transpl. 2011 Oct 27;

Authors: Gridelli B, Vizzini G, Pietrosi G, Luca A, Spada M, Gruttadauria S, Cintorino D, Amico G, Chinnici C, Miki T, Schmelzer E, Conaldi PG, Triolo F, Gerlach JC

Abstract
Background: While hepatic cell transplantation (Tx) holds the promise to bridge to whole liver transplantation for patients in endstage chronic liver failure, suitable cell populations are in debate. In addition to hepatic cells, mesenchymal stromal cells (MSC) and hematopoietic stem cells (HSC) are also considered as alternative cell sources for initial clinical cell Tx work. Fetal liver tissue contains potential progenitors of all these cell lineages. Traditional isolation techniques based on collagenase incubation of tissue fragments only yield a fraction of the available cell number. Methods and Findings: We demonstrate a five-step method using a portal vein in situ perfusion technique for tissue from the late 2nd trimester that results in high viabilities known for adult liver vascular perfusion and addresses the low cell yield of the conventional digestion methods and reduces tissue exposure to collagenase by 4-fold. We used tissue donations from gestational week 18-22, which yielded 1.8±0.7x10(9) cells with an average viability of 78%. Since Tx of HSC as well as of MSC is of interest for the treatment of hepatic failure, we confirmed phenotypically that in addition to hepatic progenitors the resulting cell preparation contains cells expressing typical MSC and HSC markers. The percentage of fetal liver cells expressing proliferation markers is 45-fold greater than that of adult hepatocytes and is comparable to that of immortalized HepG2 cells, indicating strong proliferative capacity of fetal cells. We report a human fetal liver cell transplantation case for clinical endstage chronic liver failure using the described liver cell population. The patient's MELD score improved from 15 to 10 within the first 18 months of observation. Conclusion: The human fetal liver cell isolation protocol may be of interest for further clinical translation work on the development of liver cell-based therapies. © 2011 American Association for the Study of Liver Diseases.

PMID: 22034152 [PubMed - as supplied by publisher]


Different clinical models of CD34 + cells mobilization in patients with cardiovascular disease.
Source:  NIH.gov
Monday, 29 November 1999 17:00

Different clinical models of CD34 + cells mobilization in patients with cardiovascular disease.

J Thromb Thrombolysis. 2011 Jul;32(1):1-8

Authors: Cangiano E, Cavazza C, Campo G, Valgimigli M, Francolini G, Malagutti P, Pratola C, Ferrari R

Abstract
To test the role of necrosis, ischemia or both in bone marrow cells (BMC) mobilization in patients with cardiovascular disease. We studied three groups of patients: group 1, Iatrogenic Necrosis, with pure necrosis (28 patients undergoing transcatheter radiofrequency ablation); group 2, Ischemic Necrosis (30 patients with myocardial infarction); group 3, Pure Ischemia (24 patients with unstable angina). As control groups, we studied 27 patients with stable coronary artery disease (CAD), and 20 patients without CAD undergoing angiography for valvular diseases or cardiomiopathy. CD34 + cells and cytokines were evaluated at: T(0) (baseline), 48 h and 5, 7, 10, 14 days thereafter. We observed a significant increase of CD34 + cells at T(3) and T(4) only in Iatrogenic Necrosis and Ischemic Necrosis group. The peak of mobilization was observed ten days after the necrotic event (2.8 ± 1.4 vs. 5.9 ± 1.9 in the group 1, P = 0.03; and 3 ± 1.5 vs. 5.6 ± 2 in the group 2, P = 0.04; respectively). We found a good correlation between CD34 + and vascular endothelial growth factor (VEGF) and stromal derived factor (SDF-1α) peak values (r = 0.77 and r = 0.63, respectively). At multivariable analysis, myocardial necrosis (OR 3.5, 95%CI 2.2-4.2, P < 0.01), VEGF (OR 2, 95%CI 1.1-3, P = 0.01 as above versus below median value), and SDF-1α (OR 1.6, 95%CI 1.1-2.5, P = 0.02 as above versus below median value) emerged as independent predictors of C34 + cells increase. Myocardial necrosis with simultaneous elevation of VEGF and SDF-1α causes a significant CD34 + cells mobilization in patients with cardiovascular disease.

PMID: 21197559 [PubMed - indexed for MEDLINE]


Tumor- and organ-dependent infiltration by myeloid-derived suppressor cells.
Source:  Pub Med
Monday, 29 November 1999 17:00

Tumor- and organ-dependent infiltration by myeloid-derived suppressor cells.

Int Immunopharmacol. 2011 Jul;11(7):816-26

Authors: Younos I, Donkor M, Hoke T, Dafferner A, Samson H, Westphal S, Talmadge J

Abstract
Myeloid-derived suppressor cells (MDSCs) increase during tumor growth and following cytoreductive therapy resulting in immune dysfunction and tumor escape from host control. We report organ- and tumor-specific expansion of MDSCs, differences in their molecular and membrane phenotypes and T-cell suppressive activity. A significant increase in MDSCs was observed within the spleen, peripheral blood (PB), bone marrow (BM), lungs, and livers of mice bearing orthotopic 4T1, but not CI66 mammary tumors. The PB of 4T1 TB mice had the highest frequency of MDSCs (78.6±2.1%). Similarly, the non-parenchymal cells (NPCs) in the tumor tissue, livers and lungs of 4T1 tumor-bearing (TB) mice had an increased MDSCs frequency. Studies into Gr-1 and Ly-6C staining of MDSCs revealed significant increases in CD11b+Gr-1(dull)Ly-6C(high) and CD11b+Gr-1(bright)Ly-6C(low) subsets. The frequency of MDSCs inversely correlated with the CD3+ T-cell frequency in the spleen, and blood of 4T1 TB mice and was associated with a significant decrease in splenic and NPCs IFN-γ and IL-12 transcript levels, as well as significantly increased levels of granulocyte-macrophage colony-stimulating factor (GM-CSF), stem cell factor (SCF), granulocyte colony-stimulating factor (G-CSF), interleukin-10 (IL-10), interleukin-13 (IL-13), arginase-1 (ARG-1), nitric oxide synthase (NOS-2), vascular endothelial growth factor-A (VEGF-A) transcripts. In summary, MDSCs are significantly increased not only in lymphoid organs, but also in parenchymal organs including lungs and livers of TB mice, where they may facilitate metastasis to these organ sites.

PMID: 21376153 [PubMed - indexed for MEDLINE]


Interaction between adipose tissue stromal cells and gastric cancer cells in vitro.
Source:  Pub Med
Monday, 29 November 1999 17:00

Interaction between adipose tissue stromal cells and gastric cancer cells in vitro.

Cell Tissue Res. 2011 May;344(2):287-98

Authors: Nomoto-Kojima N, Aoki S, Uchihashi K, Matsunobu A, Koike E, Ootani A, Yonemitsu N, Fujimoto K, Toda S

Abstract
Adipose tissue exists in the gastric submucosa and subserosa. Thus, adipose tissue stromal cells (ATSCs), which include mesenchymal stem cells (MSCs), seem critical for the progression of gastric cancer but their interaction with the cancer cells is unknown. We demonstrated an interaction between these cells, using immunohistochemistry, Western blot and the collagen gel invasion assay system, in which the adenocarcinoma cells (well and poorly differentiated types, MKN28 and MKN45, respectively) were cultured on a ATSC-embedded or ATSC-non-embedded gel. ATSCs promoted the expression of the growth marker, proliferation cell nuclear antigen but inhibited that of the apoptosis marker, single-stranded DNA, in the cancer cell types. ATSCs accelerated the invasion of only MKN28 into the gel and promoted the expression of mitogen-activated protein kinase (MAPK, pERK-1/2) but decreased that of the molecularly targeted protein, HER2, in the cancer cells. ATSCs did not affect the expression of the prostaglandin biosynthetic enzyme cyclooxgenase-2 (COX-2) in the cancer cells. The COX-2 inhibitor celecoxib did not affect the morphology or invasion of the cancer cells. The cancer cell types in turn promoted the display of the myofibroblast marker, α-smooth muscle actin, whereas they decreased that of some MSC markers, e.g., CD44 and CD105, in ATSCs. The data suggest that (1) ATSCs influence the progression of gastric cancer by increasing their growth/invasion and decreasing their apoptosis through MAPK activation in a COX-2-independent way; (2) ATSCs adversely affect HER2-targeted therapy; (3) the cancer cells induce the cancer-associated myofibroblast phenotype in ATSCs.

PMID: 21384185 [PubMed - indexed for MEDLINE]


Stem cells: Mesenchymal stem cells from adipose tissue could be used to deliver gene therapy to the liver.
Source:  Pub Med
Monday, 29 November 1999 17:00

Stem cells: Mesenchymal stem cells from adipose tissue could be used to deliver gene therapy to the liver.

Nat Rev Gastroenterol Hepatol. 2011 Feb;8(2):64

Authors: Barranco C

PMID: 21381245 [PubMed]


Viability and proliferation potential of adipose-derived stem cells following labeling with a positron-emitting radiotracer.
Source:  Pub Med
Monday, 29 November 1999 17:00

Viability and proliferation potential of adipose-derived stem cells following labeling with a positron-emitting radiotracer.

Eur J Nucl Med Mol Imaging. 2011 Jul;38(7):1323-34

Authors: Elhami E, Goertzen AL, Xiang B, Deng J, Stillwell C, Mzengeza S, Arora RC, Freed D, Tian G

Abstract
PURPOSE: Adipose-derived stem cells (ASCs) have promising potential in regenerative medicine and cell therapy. Our objective is to examine the biological function of the labeled stem cells following labeling with a readily available positron emission tomography (PET) tracer, (18)F-fluoro-2-deoxy-D: -glucose (FDG). In this work we characterize labeling efficiency through assessment of FDG uptake and retention by the ASCs and the effect of FDG on cell viability, proliferation, transdifferentiation, and cell function in vitro using rat ASCs.
METHODS: Samples of 10(5) ASCs (from visceral fat tissue) were labeled with concentrations of FDG (1-55 Bq/cell) in 0.75 ml culture medium. Label uptake and retention, as a function of labeling time, FDG concentration, and efflux period were measured to determine optimum cell labeling conditions. Cell viability, proliferation, DNA structure damage, cell differentiation, and other cell functions were examined. Non-labeled ASC samples were used as a control for all experimental groups. Labeled ASCs were injected via tail vein in several healthy rats and initial cell biodistribution was assessed.
RESULTS: Our results showed that FDG uptake and retention by the stem cells did not depend on FDG concentration but on labeling and efflux periods and glucose content of the labeling and efflux media. Cell viability, transdifferentiation, and cell function were not greatly affected. DNA damage due to FDG radioactivity was acute, but reversible; cells managed to repair the damage and continue with cell cycles. Over all, FDG (up to 25 Bq/cell) did not impose severe cytotoxicity in rat ASCs. Initial biodistribution of the FDG-labeled ASCs was 80% + retention in the lungs. In the delayed whole-body images (2-3 h postinjection) there was some activity distribution resembling typical FDG uptake patterns.
CONCLUSION: For in vivo cell tracking studies with PET tracers, the parameter of interest is the amount of radiotracer that is present in the cells being labeled and consequent biological effects. From our study we developed a labeling protocol for labeling ASCs with a readily available PET tracer, FDG. Our results indicate that ASCs can be safely labeled with FDG concentration up to 25 Bq/cell, without compromising their biological function. A labeling period of 90 min in glucose-free medium and efflux of 60 min in complete media resulted in optimum label retention, i.e., 60% + by the stem cells. The initial biodistribution of the implanted FDG-labeled stem cells can be monitored using microPET imaging.

PMID: 21380591 [PubMed - indexed for MEDLINE]


[Use of adipose tissue in regenerative medicine].
Source:  Pub Med
Monday, 29 November 1999 17:00

[Use of adipose tissue in regenerative medicine].

Transfus Clin Biol. 2011 Apr;18(2):124-8

Authors: Casteilla L, Planat-Benard V, Bourin P, Laharrague P, Cousin B

Abstract
Adipose tissue is abundant and well known for its involvement in obesity and associated metabolic disorders. Its uses in regenerative medicine recently attracted many investigators, as large amounts of this tissue can be easily obtained using liposuction and it contains several populations of immature cells. The largest pool of such cells corresponds to immature stromal cells, called adipose-derived stromal cells (ADSCs). These cells are purified after proteolytic digestion of adipose tissue and selection by an adherent step. ADSCs display many common features with mesenchymal stem cells derived from bone marrow, including paracrine activity, but with some specific features, among which a greater angiogenic potential. This potential is now investigating at clinical level to treat critical ischemic hindlimb by autologous cells. Other potentials are also investigated and the treatment of fistula associated or not with Crohn's disease is reaching now phase III level.

PMID: 21397545 [PubMed - indexed for MEDLINE]


Human mesenchymal stem cells are susceptible to lysis by CD8+ T-cells and NK cells.
Source:  Pub Med
Monday, 29 November 1999 17:00

Human mesenchymal stem cells are susceptible to lysis by CD8+ T-cells and NK cells.

Cell Transplant. 2011 Mar 7;

Authors: Crop MJ, Korevaar SS, de Kuiper R, Ijzermans JN, van Besouw NM, Baan CC, Weimar W, Hoogduijn MJ

Abstract
Background. There is growing interest in the use of mesenchymal stem cells (MSC) to improve the outcome of organ transplantation. The immunogenicity of MSC is, however, unclear and is important for the efficacy of MSC therapy and for potential sensitization against donor antigens. We investigated the susceptibility of autologous and allogeneic MSC for lysis by CD8+ T-lymphocytes and NK cells in a kidney transplant setting. Methods. MSC were derived from adipose tissue of human kidney donors and were CD90+, CD105+, CD166+ and HLA class I+. They showed differentiation ability and immunosuppressive capacity. Lysis of MSC by PBMC, FACS-sorted CD8+ T-cells and NK cells was measured by europium release assay. Results. Allogeneic MSC were susceptible for lysis by cytotoxic CD8+ T-cells and NK cells, while autologous MSC were lysed by NK cells only. NK cell-mediated lysis was inversely correlated with the expression of HLA class I on MSC. Lysis of autologous MSC was not dependent of culturing of MSC in FBS, and MSC in suspension as well as adherent to plastic were lysed by NK cells. Pre-transplant recipient PBMC did not lyse donor MSC, but PBMC isolated 3, 6 and 12 months after transplantation showed increasing lysing ability. After 12 months, CD8+ T-cell mediated lysis of donor MSC persisted, indicating there was no evidence for desensitization against donor MSC. Conclusions. Lysis of MSC is important to take into account when MSC are considered for clinical application. Our results suggest that the HLA background of MSC and timing of MSC administration are important for the efficacy of MSC therapy.

PMID: 21396164 [PubMed - as supplied by publisher]


Complete pulp regeneration after pulpectomy by transplantation of CD105+ stem cells with stromal cell-derived factor-1.
Source:  Pub Med
Monday, 29 November 1999 17:00

Complete pulp regeneration after pulpectomy by transplantation of CD105+ stem cells with stromal cell-derived factor-1.

Tissue Eng Part A. 2011 Aug;17(15-16):1911-20

Authors: Iohara K, Imabayashi K, Ishizaka R, Watanabe A, Nabekura J, Ito M, Matsushita K, Nakamura H, Nakashima M

Abstract
Loss of pulp due to caries and pulpitis leads to loss of teeth and reduced quality of life. Thus, there is an unmet need for regeneration of pulp. A promising approach is stem cell therapy. Autologous pulp stem/progenitor (CD105(+)) cells were transplanted into a root canal with stromal cell-derived factor-1 (SDF-1) after pulpectomy in mature teeth with complete apical closure in dogs. The root canal was successfully filled with regenerated pulp including nerves and vasculature by day 14, followed by new dentin formation along the dentinal wall. The newly regenerated tissue was significantly larger in the transplantation of pulp CD105(+) cells with SDF-1 compared with those of adipose CD105(+) cells with SDF-1 or unfractionated total pulp cells with SDF-1. The pulp CD105(+) cells highly expressed angiogenic/neurotrophic factors compared with other cells and localized in the vicinity of newly formed capillaries after transplantation, demonstrating its potent trophic effects on neovascularization. Two-dimensional electrophoretic analyses and real-time reverse transcription-polymerase chain reaction analyses demonstrated that the qualitative and quantitative protein and mRNA expression patterns of the regenerated pulp were similar to those of normal pulp. Thus, this novel stem cell therapy is the first demonstration of complete pulp regeneration, implying novel treatment to preserve and save teeth.

PMID: 21417716 [PubMed - in process]


Muscle derived stem cell contains the potential to enhance long term retention as well as an aesthetic outcome of autologous fat grafting.
Source:  Pub Med
Monday, 29 November 1999 17:00

Muscle derived stem cell contains the potential to enhance long term retention as well as an aesthetic outcome of autologous fat grafting.

Med Hypotheses. 2011 Jun;76(6):805-8

Authors: Han D, Ma Z, Zhang P, Yang JF, Zhang Y, Yang D, Liu J

Abstract
Autologous fat graft has been mentioned as a prospective source of soft-tissue filler for decades. It gives a natural consistency, is easy and safe to harvest, exhibits no hypersensitivity or foreign body reactions, and is readily available. However, the traditional fat grafting has its limitations in long term process, such as partial necrosis, loss of volume, and internal calcification. They all compromise the functional and aesthetic outcome of this procedure. In recent studies, the best results were obtained by transplanting fat tissue inside muscle, thus benefiting from its better blood supply. Muscle-derived stem cells have recently emerged as a promising source of multipotent cells which give rise to muscle fibers within muscular environment. Previous studies have also proved that muscle-derived stem cells are capable of releasing various kinds of angiogenesis agents, such as VEGF, HGF, and FGF. These cytokines are known to promote revascularization. Based on the foregoing facts, we postulate that co-transplant of autologous fat and muscle derived stem cells may enhance the long term retention and aesthetic outcome of fat grafting.

PMID: 21419577 [PubMed - indexed for MEDLINE]


Therapeutic lymphangiogenesis using stem cell and VEGF-C hydrogel.
Source:  Pub Med
Monday, 29 November 1999 17:00

Therapeutic lymphangiogenesis using stem cell and VEGF-C hydrogel.

Biomaterials. 2011 Jul;32(19):4415-23

Authors: Hwang JH, Kim IG, Lee JY, Piao S, Lee DS, Lee TS, Ra JC, Lee JY

Abstract
Lymphedema is a manifestation of lymphatic system insufficiency. It arises from primary lymphatic dysplasia or secondary obliteration after lymph node dissection or irradiation. Although improvement of swelling can be achieved by comprehensive non-operative therapy, treatment of this condition requires lifelong care and good compliance. Recently molecular-based treatments using VEGF-C have been investigated by several researchers. We designed the present study to determine whether the therapeutic efficacy of implanted human adipose-derived stem cells (hADSCs) could be improved by applying a gelatin hydrogel containing VEGF-C (VEGF-C hydrogel) to the site of tissue injury in a lymphedema mouse model. Four weeks after the operation, we evaluated edema and determined lymphatic vessel density at various post-operative time points. Mice treated with hADSCs and VEGF-C hydrogel showed a significantly decreased dermal edema depth compared to the groups of mice that received hADSCs only or VEGF-C hydrogel only. Immunohistochemical analysis also revealed that the hADSC/VEGF-C hydrogel group showed significantly greater lymphatic vessel regeneration than all the other groups. hADSCs were detected in the implantation sites of all mice in the hADSC/VEGF-C group, and exhibited a lymphatic endothelial differentiation phenotype as determined by co-staining PKH-labeled hADSCs for the lymphatic marker LYVE-1. Our results suggest that co-administration of hADSCs and VEGF-C hydrogel has a substantial positive effect on lymphangiogenesis.

PMID: 21421266 [PubMed - indexed for MEDLINE]


Controlled release of cell-permeable gene complex from poly(L-lactide) scaffold for enhanced stem cell tissue engineering.
Source:  Pub Med
Monday, 29 November 1999 17:00

Controlled release of cell-permeable gene complex from poly(L-lactide) scaffold for enhanced stem cell tissue engineering.

J Control Release. 2011 Jun 10;152(2):294-302

Authors: Jung MR, Shim IK, Kim ES, Park YJ, Yang YI, Lee SK, Lee SJ

Abstract
The use of tissue engineering to deliver genes to stem cells has been impeded by low transfection efficiency of the inserted gene and poor retention at the target site. Herein, we describe the use of non-viral gene transfer by cell-permeable peptide (CPP) to increase the transfection efficiency. The combination of this technique with the use of a controlled release concept using a poly (l-lactide) scaffold allowed for prolonged uptake in stem cells. High transfection efficiency was obtained using a human-derived arginine-rich peptide denoted as Hph-1 (YARVRRRGPRR). The formation of complex between pDNA and Hph-1 was monitored using gel retardation tests to measure size and zeta potential. Complex formation was further assessed using a DNase I protection assay. A sustained gene delivery system was developed using a fibrous 3-D scaffold coated with pDNA/Hph-1 complexes. Transfection efficiency and the mean fluorescence intensity of human adipose-derived stem cells (hASCs) on the sustained delivery scaffold were compared to those of cells transfected via bolus delivery. Plasmid DNA completely bound Hph-1 at a negative-to-positive (N/P) charge ratio of 10. After complex formation, Hph-1 appeared to effectively protect pDNA against DNase I attack and exhibited cytotoxicity markedly lower than that of the pDNA/PEI complex. Plasmid DNA/Hph-1 complexes were released from the scaffolds over 14days and were successfully transfected hASCs seeded on the scaffolds. Flow cytometry revealed that the transfection efficiency in hASCs treated with pDNA/Hph-1 complex was approximately 5-fold higher than that in cells transfected using Lipofectamine. The sustained delivery system showed a significantly higher transfection efficiency and remained able to transfect cells for a longer period of time than bolus delivery. These results suggest that cell-scaffold-based tissue regeneration can be further improved by transduction concept using CPP and controlled release using polymeric scaffold.

PMID: 21420455 [PubMed - indexed for MEDLINE]


Effects of expanded human adipose tissue-derived mesenchymal stem cells on the viability of cryopreserved fat grafts in the nude mouse.
Source:  Pub Med
Monday, 29 November 1999 17:00

Effects of expanded human adipose tissue-derived mesenchymal stem cells on the viability of cryopreserved fat grafts in the nude mouse.

Int J Med Sci. 2011;8(3):231-8

Authors: Ko MS, Jung JY, Shin IS, Choi EW, Kim JH, Kang SK, Ra JC

Abstract
Adipose-derived mesenchymal stem cells (AdMSCs) augment the ability to contribute to microvascular remodeling in vivo and to modulate vascular stability in fresh fat grafts. Although cryopreserved adipose tissue is frequently used for soft tissue augmentation, the viability of the fat graft is poor. The effects of culture-expanded human adipose tissue-derived mesenchymal stem cells (hAdMSCs) on the survival and quality of the cryopreserved fat graft were determined. hAdMSCs from the same donor were mixed with fat tissues cryopreserved at -70 °C for 8 weeks and injected subcutaneously into 6-week-old BALB/c-nu nude mice. Graft volume and weight were measured, and histology was evaluated 4 and 15 weeks post-transplantation. The hAdMSC-treated group showed significantly enhanced graft volume and weight. The histological evaluation demonstrated significantly better fat cell integrity compared with the vehicle-treated control 4 weeks post-transplantation. No significant difference in graft weight, volume, or histological parameters was found among the groups 15 weeks post-transplantation. The hAdMSCs enhanced the survival and quality of transplanted cryopreserved fat tissues. Cultured and expanded hAdMSCs have reconstructive capacity in cryopreserved fat grafting by increasing the number of stem cells.

PMID: 21448310 [PubMed - indexed for MEDLINE]


[Human mesenchymal stem cell biology].
Source:  Pub Med
Monday, 29 November 1999 17:00

[Human mesenchymal stem cell biology].

Med Sci (Paris). 2011 Mar;27(3):261-7

Authors: Charbord P, Casteilla L

Abstract
This brief overview summarises the main characteristics of bone marrow mesenchymal stem cells and of adipose-derived stem cells: methods of obtention, phenotype, differentiation potential, hematopoiesis-supportive (stromal) capacity, and immunosuppressive properties. Two points are discussed in detail: 1) criteria for stemness: multipotency, self-renewal, plasticity, and 2) the repair mechanisms implicated in the different indications of cell therapy using these cells: reconstitution of the tissue functional compartment by repopulation consequent to proliferation and differentiation or reprogrammation, stromal effects by secretion of angiogenic, anti-apoptotic, anti-fibrogenic factors, molecules involved in the regulation of inflammation, etc.

PMID: 21447295 [PubMed - indexed for MEDLINE]


Panel development for multicolor flow-cytometry testing of proliferation and immunophenotype in hMSCs.
Source:  Pub Med
Monday, 29 November 1999 17:00

Panel development for multicolor flow-cytometry testing of proliferation and immunophenotype in hMSCs.

Methods Mol Biol. 2011;698:367-85

Authors: Bradford JA, Clarke ST

Abstract
Adult human mesenchymal stem cells (hMSC) are rare fibroblast-like cells capable of differentiation into a variety of cell tissues which include bone, cartilage, muscle, ligament, tendon, and adipose. Normal adult bone marrow and adipose tissue are the most common sources of these cells. The International Society for Cellular Therapy (ISCT) has proposed a set of standards to define hMSC for laboratory investigations and preclinical studies: adherence to plastic in standard culture conditions; in vitro differentiation into osteoblasts, adipocytes, and chondroblasts; and specific surface antigen expression. Direct measurement of proliferation combined with simultaneous detection of the ISCT-consensus immunophenotypic profile provides data that is used to determine the differentiation status and health of the cells. Flow cytometry provides a powerful technology that is routinely used to simultaneously and rapidly measure multiple parameters in a single sample. This chapter describes a flow cytometric panel for the simultaneous detection of immunophenotypic profile, proliferative capacity, and DNA content measurement in hMSC. Because a relatively small number of cells are needed with this approach, measurements can be made with minimal impact on expansion potential. The ability to assess antigen expression and proliferative status enables the investigator to make informed decisions on expansion and harvesting.

PMID: 21431532 [PubMed - indexed for MEDLINE]


Phenotypic analysis and differentiation of murine mesenchymal stem cells.
Source:  Pub Med
Monday, 29 November 1999 17:00

Phenotypic analysis and differentiation of murine mesenchymal stem cells.

Methods Mol Biol. 2011;698:331-50

Authors: da Silva Meirelles L, Covas DT

Abstract
Mesenchymal stem cells (MSCs) hold great promise as therapeutic tools to treat different types of disease and the use of preclinical animal models is mandatory for the development of novel MSC-based therapies. The mouse is one of the most important species used for preclinical experiments and, by extension, so is the isolation and characterization of murine MSCs. This chapter presents methods for the phenotypic analysis of cultured murine MSCs.

PMID: 21431530 [PubMed - indexed for MEDLINE]


Osteogenic differentiation of human multipotent mesenchymal stromal cells.
Source:  Pub Med
Monday, 29 November 1999 17:00

Osteogenic differentiation of human multipotent mesenchymal stromal cells.

Methods Mol Biol. 2011;698:201-14

Authors: Gupta DM, Panetta NJ, Longaker MT

Abstract
A comprehensive knowledge of the molecular biology underlying osteogenic differentiation in a controlled, laboratory setting may promise optimization of future cell-based tissue engineering strategies for clinical problems. The scope of this review encompasses a discussion of the methodology utilized to perform such studies. Our laboratory routinely performs both in vitro and in vivo assays underlying osteogenic differentiation, and the widespread use of singular methodology across multiple investigators and institutions promises great advancements for the skeletal tissue engineering community.

PMID: 21431521 [PubMed - indexed for MEDLINE]


Short and long term fate of human AMSC subcutaneously injected in mice.
Source:  Pub Med
Monday, 29 November 1999 17:00

Short and long term fate of human AMSC subcutaneously injected in mice.

World J Stem Cells. 2011 Jun 26;3(6):53-62

Authors: López-Iglesias P, Blázquez-Martínez A, Fernández-Delgado J, Regadera J, Nistal M, Miguel MP

Abstract
AIM: To study the ability of human adipose-derived mesenchymal stem cells (AMSCs) to survive over the short and long term, their biodistribution and their biosafety in vivo in tumor-prone environments.
METHODS: We subcutaneously injected human AMSCs from different human donors into immunodeficient SCID mice over both short- (2 and 4 mo) and long- (17 mo) term in young, and aged tumor-prone mice. Presence of human cells was studied by immunohistochemistry and polymerase chain reaction analysis in all organs of injected mice.
RESULTS: Subcutaneously injected AMSCs did not form teratomas at any time point. They did not migrate but remained at the site of injection regardless of animal age, and did not fuse with host cells in any organ examined. AMSCs survived in vivo for at least 17 mo after injection, and differentiated into fibroblasts of the subdermic connective tissue and into mature adipocytes of fat tissue, exclusively at the site of injection.
CONCLUSION: Our results support the assertion that AMSC may be safe candidates for therapy when injected subcutaneously because of their long term inability to form teratomas.

PMID: 21860670 [PubMed - in process]


In vitro migration capacity of human adipose tissue-derived mesenchymal stem cells reflects their expression of receptors for chemokines and growth factors.
Source:  Pub Med
Monday, 29 November 1999 17:00

In vitro migration capacity of human adipose tissue-derived mesenchymal stem cells reflects their expression of receptors for chemokines and growth factors.

Exp Mol Med. 2011 Oct 31;43(10):596-603

Authors: Baek SJ, Kang SK, Ra JC

Abstract
The homing properties of adipose tissue-derived mesenchymal stem cells (AdMSCs) have stimulated intravenous applications for their use in stem cell therapy. However, the soluble factors and corresponding cellular receptors responsible for inducing chemotaxis of AdMSCs have not yet been reported. In the present study, the migration capacity of human AdMSCs (hAdMSCs) toward various cytokines or growth factors (GFs) and the expression of their receptors were determined. In a conventional migration assay, PDGF-AB, TGF-β1, and TNF-α showed the most effective chemoattractant activity. When AdMSCs were preincubated with various chemokines or GF, and then allowed to migrate toward medium containing 10% FBS, those preincubated with TNF-α showed the highest migratory activity. Next, hAdMSCs were either preincubated or not with TNF-α, and allowed to migrate in response to various GFs or chemokines. Prestimulation with TNF-α increased the migration activity of hAdMSCs compared to unstimulated hAdMSCs. When analyzed by FACS and RT-PCR methods, hAdMSCs were found to express C-C chemokine receptor type 1 (CCR1), CCR7, C-X-C chemokine receptor type 4 (CXCR4), CXCR5, CXCR6, EGF receptor, fibroblast growth factor receptor 1, TGF-β receptor 2, TNF receptor superfamily member 1A, PDGF receptor A and PDGF receptor B at both the protein and the mRNA levels. These results indicate that the migration capacity of hAdMSCs is controlled by various GFs and chemokines. Prior in vitro modulation of the homing capacity of hAdMSCs could stimulate their movement into injured sites in vivo when administered intravenously, thereby improving their therapeutic potential.

PMID: 21847008 [PubMed - in process]


[Expression of human bone morphogenetic protein 7 gene in adipose-derived stem cells and its effects on osteogenic phenotype].
Source:  Pub Med
Monday, 29 November 1999 17:00

[Expression of human bone morphogenetic protein 7 gene in adipose-derived stem cells and its effects on osteogenic phenotype].

Zhongguo Xiu Fu Chong Jian Wai Ke Za Zhi. 2011 Jul;25(7):848-53

Authors: Ren Y, Han C, Jia Y, Yin H, Li S

Abstract
OBJECTIVE: To construct the eukaryotic expression vector of human bone morphogenetic protein 7 (hBMP-7) gene so as to observe its expression in rabbit adipose-derived stem cells (ADSCs) and its effects on osteogenic phenotype.
METHODS: Several healthy 3-month-old Japanese rabbits of clean grade were chosen, female or male and weighing 3-4 kg. ADSCs were isolated and cultured with collagenase digestion, then were detected and identified by CD44, CD49d, and CD106 immunofluorescence staining. The eukaryotic expression vector of hBMP-7 gene (pcDNA3.1-hBMP-7) was constructed, which was transfected into rabbit ADSCs (3rd passage) by Lipofectamine 2000 after identified, then the expression of hBMP-7 in transfected ADSCs was detected. The alkaline phosphatase (ALP) level and the collagen type I expression were detected by intracellular ALP spectrophotometry and immunofluorescence, respectively to assess the effect of hBMP-7 gene on the osteoblastic differentiation of ADSCs.
RESULTS: ADSCs mostly presented fusiform and polygon shape with positive expressions of CD44 and CD49d and negative expression of CD106. The eukaryotic expression vector of pcDNA3.1-hBMP-7 gene was successfully constructed and the expression of hBMP-7 was confirmed in ADSCs by immunohistochemical staining. The intracellular ALP quantitative detection showed that the activity of ALP was significantly higher in pcNDA3.1-hBMP-7 transfected group (experimental group) than in pcDNA3.1 transfected group (control group) at 7, 10, and 14 days after transfection (P < 0.05). The expression of collagen type I was higher in experimental group than in control group at 7 and 14 days after transfection (P < 0.05).
CONCLUSION: The eukaryotic expression vector of pcDNA3.1-hBMP-7 gene is successfully constructed, which can express in ADSCs. The expressions of collagen type I and ALP in experimental group are higher than those in control group, which lays a basis for the local gene therapy of skeletal regeneration.

PMID: 21818953 [PubMed - in process]


HMG-CoA reductase inhibitor augments the serum total cholesterol-lowering effect of human adipose tissue-derived multilineage progenitor cells in hyperlipidemic homozygous Watanabe rabbits.
Source:  Pub Med
Monday, 29 November 1999 17:00

HMG-CoA reductase inhibitor augments the serum total cholesterol-lowering effect of human adipose tissue-derived multilineage progenitor cells in hyperlipidemic homozygous Watanabe rabbits.

Biochem Biophys Res Commun. 2011 Aug 19;412(1):50-4

Authors: Saga A, Okura H, Soeda M, Tani J, Fumimoto Y, Komoda H, Moriyama M, Moriyama H, Yamashita S, Ichinose A, Daimon T, Hayakawa T, Matsuyama A

Abstract
Familial hypercholesterolemia (FH) is an autosomal codominant disease characterized by high concentrations of proatherogenic lipoproteins secondary to deficiency in low-density lipoprotein (LDL) receptor. We reported recently the use of in situ stem cell therapy of human adipose tissue-derived multilineage progenitor cells (hADMPCs) in lowering serum total cholesterol in the homozygous Watanabe heritable hyperlipidemic (WHHL) rabbits, an animal model of homozygous FH. Here we demonstrate that pravastatin, an HMG-CoA reductase inhibitor, augmented the cholesterol-lowering effect of transplanted hADMPCs and enhanced LDL clearance in homozygous WHHL rabbit. The results suggest the potential beneficial effects of in situ stem cell therapy in concert with appropriately selected pharmaceutical agents, in regenerative medicine.

PMID: 21806964 [PubMed - indexed for MEDLINE]


[Outgrowth of neuronal axons on adipose-derived stem cell transplanting for treatment of cerebral infarction in rats].
Source:  Pub Med
Monday, 29 November 1999 17:00

[Outgrowth of neuronal axons on adipose-derived stem cell transplanting for treatment of cerebral infarction in rats].

Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi. 2011 Aug;27(8):868-71

Authors: Chen AZ, Liu N, Huang H, Lin FF, Liu DS, Lin XH

Abstract
AIM: To investigate the effects of adipose-derived stem cell (ADSC) transplanting on the outgrowth of neuronal axons and the expressions of GFAP, Neuritin, NF-200 in the brain post focal cerebral ischemia in rats.
METHODS: 54 male adult Sprague-Dawley rats were randomly divided into 3 groups: sham-operated group, middle cerebral artery occlusion (MCAO) group and MCAO+ADSC-treated group (n=18 in each group). A permenant focal cerebal ischemia model was established using modified Longa's method ADSC was labeled by DAPI before the transplantation. One day after MCAO, 30 μL of cell suspension containing 1×10(6); cells were injected into the lateral ventricle of MCAO+ADSC-treated group. At 7 d, 14 d and 28 d after MCAO, the expressions of GFAP, Neuritin and NF-200 were detected in ischemic region by Western blot and Immunofluorescence analysis.
RESULTS: DAPI staining positive cells were observed around the cerebral infarcted area in the ADSC group. The expressions of Neuritin, NF200 were higher, but GFAP was lower than that of the MCAO group at 7 d, 14 d and 28 d (P<0.05).
CONCLUSION: The transplantation of ADSC can induce regeneration and repairment of impaired neuronal axons in rat brain after cerebral ischemia, partly by inhibiting the expression of GFAP and enhancing the expressions of Neuritin, NF-200 in the brain.

PMID: 21806883 [PubMed - indexed for MEDLINE]


Therapeutic effect of osteogenically induced adipose derived stem cells on vascular deprivation-induced osteonecrosis of the femoral head in rabbits.
Source:  Pub Med
Monday, 29 November 1999 17:00

Therapeutic effect of osteogenically induced adipose derived stem cells on vascular deprivation-induced osteonecrosis of the femoral head in rabbits.

Chin J Traumatol. 2011 Aug 1;14(4):215-20

Authors: Aimaiti A, Saiwulaiti Y, Saiyiti M, Wang YH, Cui L, Yusufu A

Abstract
Objective: To explore the therapeutic effect of osteogenically induced adipose-derived stem cells (ADSCs) on vascular deprivation-induced osteonecrosis of the femoral head (ONFH) in rabbit model. Methods: Vascular deprivation-induced ONFH was established by intramuscular injection of methylpre- dnisolone, and vascular occlusion of the capital femoral epiphysis by electrocoagulation in adult New Zealand white rabbits. Eight weeks after the establishment of vascular deprivation-induced ONFH, animals were randomly divided into three equal groups. In Group A (control), no therapy was given. In Group B, core decompression was performed by drilling a hole (1.2 mm in diameter) from the outer cortex 2.5 cm distal to the proximal end of the greater trochanter. In Group C, 1 multiply 10(7) osteogenically induced ADSCs were resuspended in 0.5 ml PBS, and then injected directly into the femoral head. Femoral head specimens were obtained at postoperative 8 weeks. The bone formation and three-dimensional microstructure of the femoral head was evaluated by micro-computed tomography scans. Immunohistochemical analysis was performed to detect the expression of osteocalcin. Angiogenesis and repair of the femoral head were observed histologically.Results: In trabecular bone at the proximal femur region, the trabecular volume was higher in Group C (130.70 mm(3)+/-4.33 mm(3)) than that in Groups A (101.07 mm(3)+/-7.76 mm(3)) and B (107.89 mm(3)+/-8.68 mm(3), P less than 0.01). Bone volume was significantly increased in Group C (40.09 mm(3)+/-6.35 mm(3)) than in Groups A (29.65 mm(3)+/-4.61 mm(3)) and B (31.80 mm(3)+/-4.01 mm(3), P less than 0.01). The trabecular number was higher in Groups C (1.58+/-0.25) than other two groups (1.15+/-0.18, 1.16+/-0.21, P less than 0.01). Bone mineral density showed statistically significant difference between Groups C and A or B (375.38+/-23.06) mg HA/ccm, vs (313.73+/-19.30) mg HA/ccm and (316.09+/-16.45) mg HA/ccm, P less than 0.01). Histological examination indicated that there was more new bone formation in Group C than in other groups.Conclusion: Treatment with autologous osteogeni-cally induced ADSCs transplantation results in an enhanced osteogenesis and microstructure of the vascular deprivation-induced osteonecrosis in rabbits.

PMID: 21801665 [PubMed - in process]


Tracking intracavernously injected adipose-derived stem cells to bone marrow.
Source:  Pub Med
Monday, 29 November 1999 17:00

Tracking intracavernously injected adipose-derived stem cells to bone marrow.

Int J Impot Res. 2011 Jul 28;

Authors: Lin G, Qiu X, Fandel T, Banie L, Wang G, Lue TF, Lin CS

Abstract
The intracavernous (i.c.) injection of stem cells (SCs) has been shown to improve erectile function in various erectile dysfunction (ED) animal models. However, the tissue distribution of the injected cells remains unknown. In this study we tracked i.c.-injected adipose-derived stem cells (ADSCs) in various tissues. Rat paratesticular fat was processed for ADSC isolation and culture. The animals were then subject to cavernous nerve (CN) crush injury or sham operation, followed by i.c. injection of 1 million autologous or allogeneic ADSCs that were labeled with 5-ethynyl-2-deoxyuridine (EdU). Another group of rats received i.c. injection of EdU-labeled allogeneic penile smooth muscle cells (PSMCs). At 2 and 7 days post injection, penises and femoral bone marrow were processed for histological analyses. Whole femoral bone marrows were also analyzed for EdU-positive cells by flow cytometry. The results show that ADSCs exited the penis within days of i.c. injection and migrated preferentially to bone marrow. Allogenicity did not affect the bone marrow appearance of ADSCs at either 2 or 7 days, whereas CN injury reduced the number of ADSCs in bone marrow significantly at 7 but not 2 days. The significance of these results in relation to SC therapy for ED is discussed.International Journal of Impotence Research advance online publication, 28 July 2011; doi:10.1038/ijir.2011.38.

PMID: 21796145 [PubMed - as supplied by publisher]


Stem Cell Therapy for Erectile Dysfunction: A Critical Review.
Source:  Pub Med
Monday, 29 November 1999 17:00

Stem Cell Therapy for Erectile Dysfunction: A Critical Review.

Stem Cells Dev. 2011 Sep 7;

Authors: Lin CS, Xin ZC, Wang Z, Deng C, Huang YC, Lin G, Lue TF

Abstract
Erectile dysfunction (ED) is a prevailing health problem that seriously impacts quality of life. Current treatment options are less effective for patients having cavernous nerve (CN) injury or diabetes mellitus-related ED. These 2 types of ED are thus the main focus of past and current stem cell (SC) therapy studies. In a total of 16 studies so far, rats were exclusively used as disease models and SCs were mostly derived from bone marrow, adipose tissue, or skeletal muscle. For tracking, SCs were labeled with LacZ, green fluorescent protein, 4',6-diamidino-2-phenylindole, DiI, bromodeoxyuridine, or 5-ethynyl-2-deoxyuridine, some of which might have led to data misinterpretation. SC transplantation was done exclusively by intracavernous (IC) injection, which has been recently shown to have systemic effects. Functional assessment was done exclusively by measuring increases of IC pressure during electrostimulation of CN. Histological assessment usually focused on endothelial, smooth muscle, and CN contents in the penis. In general, favorable outcomes have been obtained in all trials so far, although whether SCs had differentiated into specific cell lineages remains controversial. Recent studies have shown that intracavernously injected SCs rapidly escaped the penis and homed into bone marrow. This could perhaps explain why intracavernously injected SCs had systemic antidiabetic effects and prolonged anti-ED effects. These hypotheses and the differentiation-versus-paracrine debate require further investigation.

PMID: 21793654 [PubMed - as supplied by publisher]


The effect of human adipose-derived stem cells on healing of ischemic wounds in a diabetic nude mouse model.
Source:  Pub Med
Monday, 29 November 1999 17:00

The effect of human adipose-derived stem cells on healing of ischemic wounds in a diabetic nude mouse model.

Plast Reconstr Surg. 2011 Aug;128(2):387-94

Authors: Kim EK, Li G, Lee TJ, Hong JP

Abstract
BACKGROUND: Among hundreds of pathophysiologic factors that impair healing of the diabetic foot, diminished perfusion and neovascularization are critical. This study aimed to show the effect of human adipose-derived stem cells on healing of ischemic wounds in diabetic nude mice and thus to estimate the possibilities of adipose-derived stem cells for diabetic wound care.
METHODS: Sixty nude mice were assigned randomly into group Ia (diabetic control, n = 20), group Ib (diabetic experimental, n = 20), or group II (nondiabetic control, n = 20). After creation of an ischemic limb, human adipose-derived stem cells were injected locally. Gross and histologic observations were made after 3, 7, and 25 days, and plasma and tissue levels of vascular endothelial growth factor were quantified.
RESULTS: Group Ia animals typically showed gangrene formation with a delayed and sustained inflammatory reaction, which led to a high rate of autoamputation and a lower survival rate. Group Ib animals had a tendency for earlier and abundant neovessel formation and better tissue remodeling rather than fibrotic cicatrization, resulting in lower rate of autoamputation and a survival rate comparable to group II. Surviving stem cells were identified at day 25, but no specific differentiation was observed. Plasma and tissue vascular endothelial growth factor level in group Ib was higher than in group Ia and comparable to that in group II.
CONCLUSIONS: This study demonstrated the beneficial effect of human adipose-derived stem cells on healing of ischemic wound in diabetic nude mice. Elevation of vascular endothelial growth factor levels in plasma and tissue suggests the importance of secretory factor, which regulates local angiogenesis and triggers a systemic response.

PMID: 21788830 [PubMed - indexed for MEDLINE]


Autologous transplantation of adipose-derived mesenchymal stem cells markedly reduced acute ischemia-reperfusion lung injury in a rodent model.
Source:  Pub Med
Monday, 29 November 1999 17:00

Autologous transplantation of adipose-derived mesenchymal stem cells markedly reduced acute ischemia-reperfusion lung injury in a rodent model.

J Transl Med. 2011;9:118

Authors: Sun CK, Yen CH, Lin YC, Tsai TH, Chang LT, Kao YH, Chua S, Fu M, Ko SF, Leu S, Yip HK

Abstract
BACKGROUND: This study tested the hypothesis that autologous transplantation of adipose-derived mesenchymal stem cells (ADMSCs) can effectively attenuate acute pulmonary ischemia-reperfusion (IR) injury.
METHODS: Adult male Sprague-Dawley (SD) rats (n = 24) were equally randomized into group 1 (sham control), group 2 (IR plus culture medium only), and group 3 (IR plus intravenous transplantation of 1.5 × 106 autologous ADMSCs at 1h, 6h, and 24h following IR injury). The duration of ischemia was 30 minutes, followed by 72 hours of reperfusion prior to sacrificing the animals. Blood samples were collected and lungs were harvested for analysis.
RESULTS: Blood gas analysis showed that oxygen saturation (%) was remarkably lower, whereas right ventricular systolic pressure was notably higher in group 2 than in group 3 (all p < 0.03). Histological scoring of lung parenchymal damage was notably higher in group 2 than in group 3 (all p < 0.001). Real time-PCR demonstrated remarkably higher expressions of oxidative stress, as well as inflammatory and apoptotic biomarkers in group 2 compared with group 3 (all p < 0.005). Western blot showed that vascular cell adhesion molecule (VCAM)-1, intercellular adhesion molecule (ICAM)-1, oxidative stress, tumor necrosis factor-α and nuclear factor-κB were remarkably higher, whereas NAD(P)H quinone oxidoreductase 1 and heme oxygenase-1 activities were lower in group 2 compared to those in group 3 (all p < 0.004). Immunofluorescent staining demonstrated notably higher number of CD68+ cells, but significantly fewer CD31+ and vWF+ cells in group 2 than in group 3.
CONCLUSION: ADMSC therapy minimized lung damage after IR injury in a rodent model through suppressing oxidative stress and inflammatory reaction.

PMID: 21781312 [PubMed - in process]


A cell-free extract from human adipose stem cells protects mice against epilepsy.
Source:  Pub Med
Monday, 29 November 1999 17:00

A cell-free extract from human adipose stem cells protects mice against epilepsy.

Epilepsia. 2011 Sep;52(9):1617-26

Authors: Jeon D, Chu K, Lee ST, Jung KH, Kang KM, Ban JJ, Kim S, Seo JS, Won CH, Kim M, Lee SK, Roh JK

Abstract
PURPOSE: Stem cell-based therapies are being considered for various neurologic diseases, such as epilepsy. Recent studies have suggested that some effects of transplanted stem cells are due to bystander effects that modulate the host environment, rather than direct effects of cell replacement. The extract from human adipose stem cells (ASCs) that secrete multiple growth factors including cytokines and chemokines may be a potential source of bystander effects for the treatment of epilepsy, in which inflammation is thought to play an important role. Here, we investigated the effects of a cytosolic extract of human ASCs (ASCs-E) in a mouse model of epilepsy.
METHODS: Human ASCs-E, boiled ASCs-E, or fibroblast-extract (fibroblast-E) was intraperitoneally administrated to C57BL/6 mice 15 min before pilocarpine-induced status epilepticus (SE) or during chronic epileptic stage. Blood-brain barrier (BBB) leakage was evaluated by measuring Evans blue dye extravasation. Spontaneous recurrent seizure (SRS) was investigated by long-term video-electroencephalography (EEG) monitoring. The mice performed elevated plus maze, open-field, light/dark transition, and novel object recognition tasks.
KEY FINDINGS: Acute application of human ASCs-E before SE led to earlier attenuation of seizure spike activities after treatment with diazepam, reduction of BBB leakage, and inhibition of the development of epilepsy. Human ASCs-E treatment (for 7 days) during the chronic epileptic stage suppressed SRS and reduced abnormal epileptic behavioral phenotypes. However, neither boiled ASCs-E nor fibroblast-E had any effects in the experimental epilepsy model.
SIGNIFICANCE: Our results demonstrate that human ASCs-E prevents or inhibits epileptogenesis and SRS in mice. They also suggest a stem cell-based, noninvasive therapy for the treatment of epilepsy.

PMID: 21777228 [PubMed - indexed for MEDLINE]


Stem cell based cancer gene therapy.
Source:  Pub Med
Monday, 29 November 1999 17:00

Stem cell based cancer gene therapy.

Mol Pharm. 2011 Oct 3;8(5):1480-7

Authors: Cihova M, Altanerova V, Altaner C

Abstract
The attractiveness of prodrug cancer gene therapy by stem cells targeted to tumors lies in activating the prodrug directly within the tumor mass, thus avoiding systemic toxicity. Suicide gene therapy using genetically engineered mesenchymal stem cells has the advantage of being safe, because prodrug administration not only eliminates tumor cells but consequently kills the more resistant therapeutic stem cells as well. This review provides an explanation of the stem cell-targeted prodrug cancer gene therapy principle, with focus on the choice of prodrug, properties of bone marrow and adipose tissue-derived mesenchymal stem and neural stem cells as well as the mechanisms of their tumor homing ability. Therapeutic achievements of the cytosine deaminase/5-fluorocytosine prodrug system and Herpes simplex virus thymidine kinase/ganciclovir are discussed. In addition, delivery of immunostimulatory cytokines, apoptosis inducing genes, nanoparticles and antiangiogenic proteins by stem cells to tumors and metastases is discussed as a promising approach for antitumor therapy. Combinations of traditional, targeted and stem cell-directed gene therapy could significantly advance the treatment of cancer.

PMID: 21755953 [PubMed - in process]


Adipose tissue and bone marrow-derived stem cells react similarly in an ischaemia-like microenvironment.
Source:  Pub Med
Monday, 29 November 1999 17:00

Adipose tissue and bone marrow-derived stem cells react similarly in an ischaemia-like microenvironment.

J Tissue Eng Regen Med. 2011 Jul 13;

Authors: Tse KH, Kingham PJ, Novikov LN, Wiberg M

Abstract
Mesenchymal stem cells (MSCs) from adipose tissue and bone marrow are promising cell sources for autologous cell therapy of nerve injuries, as demonstrated by their intrinsic neurotrophic potential. However, extensive death of transplanted cells limits their full benefits. This study investigated the effects of ischaemia (metabolically induced by sodium azide and 2-deoxyglucose) and serum-derived mitogens on the viability and functional profile of MSCs in vitro. MSCs were more susceptible to combined, rather than individual, blockade of glycolysis and oxidative phosphorylation. Apoptosis and autophagy were involved in ischaemia-induced cell death. Chemical ischaemia alone and serum withdrawal alone induced a similar amount of cell death, with significantly different intracellular ATP maintenance. Combined ischaemia and serum deprivation had additive effects on cell death. Expression of the extracellular matrix (ECM) molecules laminin and fibronectin was attenuated under ischaemia and independent of serum level; however, BDNF and NGF levels remained relatively constant. Strong upregulation of VEGF and to a lesser extent angiopoietin-1 was observed under ischaemia but not in serum withdrawal conditions. Importantly, this study demonstrated similar reactions of MSCs derived from adipose and bone marrow tissue, in ischaemia-like and mitogen-deprived microenvironments in terms of viability, cellular energetics, cell death mechanisms and expression levels of various growth-promoting molecules. Also, the results suggest that ischaemia has a larger impact on the ability of MSCs to survive transplantation than withdrawal of mitogens. Copyright © 2011 John Wiley & Sons, Ltd.

PMID: 21751425 [PubMed - as supplied by publisher]


Effects of Human Adipose-derived Stem Cells on Cutaneous Wound Healing in Nude Mice.
Source:  Pub Med
Monday, 29 November 1999 17:00

Effects of Human Adipose-derived Stem Cells on Cutaneous Wound Healing in Nude Mice.

Ann Dermatol. 2011 May;23(2):150-5

Authors: Lee SH, Lee JH, Cho KH

Abstract
BACKGROUND: Despite numerous treatments available for deteriorated cutaneous wound healing such as a diabetic foot, there is still the need for more effective therapy. Adipose-derived stem cells (ASCs) are mesenchymal stem cells, which are self-renewing and multipotent. Mesenchymal stem cells have the potential for tissue repair and regeneration.
OBJECTIVE: To investigate the effects of human ASCs on the healing of cutaneous wounds in nude mice.
METHODS: 15-mm round full-thickness skin defects were generated on the back of BALB/c nude mice. The mice were divided into three groups for wound coverage: (i) human ASCs-populated collagen gel, (ii) human dermal fibroblasts-populated collagen gel, and (iii) collagen gel alone. Wound contraction was prevented with a splint method. Wound size was measured 10 days after injury. At 28 days histological analysis was performed.
RESULTS: Both ASCs and dermal fibroblasts accelerated wound closure, but dermal fibroblasts were more effective than ASCs. At 28 days, the dermal portion of ASCs or dermal fibroblasts wound scars were thicker than collagen gel wound scars.
CONCLUSION: ASCs and dermal fibroblasts stimulate cutaneous wound healing and improve scar thickness.

PMID: 21747612 [PubMed]


Effective myotube formation in human adipose tissue-derived stem cells expressing dystrophin and myosin heavy chain by cellular fusion with mouse C2C12 myoblasts.
Source:  Pub Med
Monday, 29 November 1999 17:00

Effective myotube formation in human adipose tissue-derived stem cells expressing dystrophin and myosin heavy chain by cellular fusion with mouse C2C12 myoblasts.

Biochem Biophys Res Commun. 2011 Apr 29;408(1):167-73

Authors: Eom YW, Lee JE, Yang MS, Jang IK, Kim HE, Lee DH, Kim YJ, Park WJ, Kong JH, Shim KY, Lee JI, Kim HS

Abstract
Stem cell therapy for muscular dystrophies requires stem cells that are able to participate in the formation of new muscle fibers. However, the differentiation steps that are the most critical for this process are not clear. We investigated the myogenic phases of human adipose tissue-derived stem cells (hASCs) step by step and the capability of myotube formation according to the differentiation phase by cellular fusion with mouse myoblast C2C12 cells. In hASCs treated with 5-azacytidine and fibroblast growth factor-2 (FGF-2) for 1 day, the early differentiation step to express MyoD and myogenin was induced by FGF-2 treatment for 6 days. Dystrophin and myosin heavy chain (MyHC) expression was induced by hASC conditioned medium in the late differentiation step. Myotubes were observed only in hASCs undergoing the late differentiation step by cellular fusion with C2C12 cells. In contrast, hASCs that were normal or in the early stage were not involved in myotube formation. Our results indicate that stem cells expressing dystrophin and MyHC are more suitable for myotube formation by co-culture with myoblasts than normal or early differentiated stem cells expressing MyoD and myogenin.

PMID: 21473854 [PubMed - indexed for MEDLINE]


Wound microenvironment sequesters adipose-derived stem cells in a murine model of reconstructive surgery in the setting of concurrent distant malignancy.
Source:  Pub Med
Monday, 29 November 1999 17:00

Wound microenvironment sequesters adipose-derived stem cells in a murine model of reconstructive surgery in the setting of concurrent distant malignancy.

Plast Reconstr Surg. 2011 Apr;127(4):1467-77

Authors: Altman AM, Prantl L, Muehlberg FL, Song YH, Seidensticker M, Butler CE, Alt EU

Abstract
BACKGROUND: It is unclear whether mesenchymal stem cells that are applied to regenerate wound tissues can migrate to existing tumors and enhance their growth. The authors investigated whether adipose-derived stem cells had any effect on the growth and progression of distant tumors when applied to a skin wound.
METHODS: The authors subcutaneously injected murine 4T1 breast cancer cells into all BALB/c-nu/nu mice. After tumor injection, mice were randomized to five groups (five mice per group) based on the means of co-introduction of green fluorescent protein-labeled adipose-derived stem cells, if any. In group 1, adipose-derived stem cells were combined and co-injected subcutaneously. In group 2, they were injected subcutaneously at a distant anatomical site. In group 3, they were injected intravenously. In group 4, they were delivered via a human acellular dermal matrix to a distant skin wound. In group 5, no adipose-derived stem cells were introduced.
RESULTS: After 2 weeks, tumor volume increased in group 1 (356.5 ± 44.4 mm(3)), followed by group 3 (256.6 ± 47.1 mm(3)) and then group 2 (201.6 ± 28.6 mm(3)). In group 4, in which adipose-derived stem cells carried on acellular dermal matrix were applied to a wound distant to the primary tumor, the tumor volume was 143.8 ± 50.9 mm(3), which was similar to that observed in the control group (group 5; 167.8 ± 29.9 mm(3)).
CONCLUSIONS: The authors' findings suggest that the wound microenvironment can retain adipose-derived stem cells, preventing their homing and stromal contribution to a distant neoplastic focus. These findings are an important first step in establishing the feasibility and safety of utilizing adipose-derived stem cell therapy for reconstructive surgery in the setting of malignancy.

PMID: 21460655 [PubMed - indexed for MEDLINE]


High-mobility group protein HMGA2-derived fragments stimulate the proliferation of chondrocytes and adipose tissue-derived stem cells.
Source:  Pub Med
Monday, 29 November 1999 17:00

High-mobility group protein HMGA2-derived fragments stimulate the proliferation of chondrocytes and adipose tissue-derived stem cells.

Eur Cell Mater. 2011;21:355-63

Authors: Richter A, Lübbing M, Frank HG, Nolte I, Bullerdiek JC, von Ahsen I

Abstract
In previous research, it was shown that recombinant HMGA2 protein enhances the proliferation of porcine chondrocytes grown in vitro, opening up promising applications of this embryonic architectural transcription factor for tissue engineering, such as in cartilage repair. In this paper, we describe the development and analyses of two synthetic fragments comprising the functional AT-hook motifs of the HMGA2 protein, as well as the nuclear transport domain. They can be synthesised up to large scales, while eliminating some of the problems of recombinant protein production, including unwanted modification or contamination by the expression hosts, or of gene therapy approaches such as uncontrolled viral integration and transgene expression even after therapy. Application of one of these peptides onto porcine hyaline cartilage chondrocytes, grown in in vitro monolayer cell culture, showed a growth-promoting effect similar to that of the wild type HMGA2 protein. Furthermore, it also promoted cell growth of adult adipose tissue derived stem cells. Due to its proliferation inducing function and vast availability, this peptide is thus suitable for further application and investigation in various fields such as tissue engineering and stem cell research.

PMID: 21484705 [PubMed - indexed for MEDLINE]


microRNAs in the regulation of adipogenesis and obesity.
Source:  Pub Med
Monday, 29 November 1999 17:00

microRNAs in the regulation of adipogenesis and obesity.

Curr Mol Med. 2011 Jun;11(4):304-16

Authors: McGregor RA, Choi MS

Abstract
Worldwide obesity is a growing health problem, associated with increased risk of chronic disease. Understanding the molecular basis of adipogenesis and fat cell development in obesity is essential to identify new biomarkers and therapeutic targets for the development of anti-obesity drugs. microRNAs (miRNAs) appear to play regulatory roles in many biological processes associated with obesity, including adipocyte differentiation, insulin action and fat metabolism. Recent studies show miRNAs are dysregulated in obese adipose tissue. During adipogenesis miRNAs can accelerate or inhibit adipocyte differentiation and hence regulate fat cell development. In addition miRNAs may regulate adipogenic lineage commitment in multipotent stem cells and hence govern fat cell numbers. Recent findings suggest miR-519d may be associated with human obesity, but larger case-control studies are needed. Few miRNA targets have been experimentally validated in adipocytes but interestingly both miR-27 and miR-519d target PPAR family members, which are well established regulators of fat cell development. In this review recent advances in our understanding of the role of miRNAs in fat cell development and obesity are discussed. The potential of miRNA based therapeutics targeting obesity is highlighted as well as recommendations for future research which could lead to a breakthrough in the treatment of obesity.

PMID: 21506921 [PubMed - indexed for MEDLINE]


Cavernous nerve repair with allogenic adipose matrix and autologous adipose-derived stem cells.
Source:  Pub Med
Monday, 29 November 1999 17:00

Cavernous nerve repair with allogenic adipose matrix and autologous adipose-derived stem cells.

Urology. 2011 Jun;77(6):1509.e1-8

Authors: Lin G, Albersen M, Harraz AM, Fandel TM, Garcia M, McGrath MH, Konety BR, Lue TF, Lin CS

Abstract
OBJECTIVES: To investigate whether adipose-derived matrix seeded with adipose-derived stem cells (ADSC) can facilitate the repair of injured cavernous nerves (CNs).
METHODS: Human and rat adipose tissues were decellularized and fabricated into various forms, including adipose tissue-derived acellular matrix thread (ADMT). ADMT seeded with ADSC were transplanted into subcutaneous space and examined for signs of inflammation. ADSC-seeded ADMTs were then used to repair CN injury in rats, followed by assessment of histology and erectile function.
RESULTS: Adipose tissue can be fabricated into acellular matrices of various shapes and sizes, including threads and sheets. Seeding of ADMT occurred rapidly: within 24 hours, 55% of the surface was covered with ADSC and within 1 week, 90% was covered. Transplantation of the seeded ADMT into the subcutaneous space of an allogenic host showed no signs of inflammatory reaction. At 3 months after grafting into CN injury rats, approximately twice as many cells were found on seeded ADMT as on unseeded ADMT. The seeded ADMT also had various degrees of S100 and neuronal nitric oxide synthase expression, suggesting CN axonal ingrowth. Rats grafted with seeded ADMT overall had the best erectile function recovery when compared with those grafted with unseeded ADMT and those ungrafted. However, as a result of large variations, the differences did not reach statistic significance (P = .07).
CONCLUSIONS: Grafting of ADSC-seeded matrix resulted in a substantial recovery of erectile function and improvement of histology. However, further refinement of the matrix architecture is needed to improve the success rate.

PMID: 21492917 [PubMed - indexed for MEDLINE]


Staged Stem Cell-enriched Tissue (SET) Injections for Soft Tissue Augmentation in Hostile Recipient Areas: A Preliminary Report.
Source:  Pub Med
Monday, 29 November 1999 17:00

Staged Stem Cell-enriched Tissue (SET) Injections for Soft Tissue Augmentation in Hostile Recipient Areas: A Preliminary Report.

Aesthetic Plast Surg. 2011 Apr 13;

Authors: Tiryaki T, Findikli N, Tiryaki D

Abstract
BACKGROUND: Autologous fat transplantation is frequently used for a variety of cosmetic treatments and difficult reconstructive indications such as involutional disorders, hemifacial atrophy, sequelae of radiation therapy, or similar problems. However, the limitations of fat transplantation are well known in such difficult cases, particularly the long-term unpredictability of volume maintenance. The ideal method of preparing autologous fat grafts optimizes tissue survival and reduces the variability of outcomes. We propose that enriching traditionally prepared fat grafts with adipose-derived regenerative cells (ADRCs) represents one such method. METHODS: Using a staged approach, we performed cell-enriched fat transfer by injecting autologous ADRCs into soft tissue that was recently grafted using traditional methods of fat transfer. Over a 3-year period, data were prospectively collected from 29 patients who underwent a single session of stem cell-enriched tissue injections (SET). RESULTS: Cell-enriched grafts ranged in volume from 10 to 390 cc per recipient area and were obtained by manual or automated processes. The mean follow-up period was 10 months. Postoperative atrophy of the injected tissue was minimal and subjectively did not change after 8 weeks. Of note, historically reported rates of atrophy range from 20 to 80%. All patients were satisfied with the primary result with no need for a secondary session except for the cosmetic cases. CONCLUSION: These preliminary results suggest that SET is safe and may provide superior results compared to traditional fat grafting. By performing the procedure in a staged approach, operating room expenses are minimized, which ultimately decreases the cost of the procedure. Adipose-derived regenerative cells may mitigate early ischemia by increasing angiogenesis, decreasing apoptosis, and modulating the local inflammatory response. This technique may be of particular value to the surgeon when grafting high volumes of fat or when faced with hostile recipient area conditions, including fibrosis and post radiation.

PMID: 21487913 [PubMed - as supplied by publisher]


Directed differentiation of motor neuron cell-like cells from human adipose-derived stem cells in vitro.
Source:  Pub Med
Monday, 29 November 1999 17:00

Directed differentiation of motor neuron cell-like cells from human adipose-derived stem cells in vitro.

Neuroreport. 2011 Jun 11;22(8):370-3

Authors: Liqing Y, Jia G, Jiqing C, Ran G, Fei C, Jie K, Yanyun W, Cheng Z

Abstract
The capacity of human adipose-derived stem cells (hADSCs) to differentiate into motor neurons and the identity of molecular factors that confer hADSCs with the competence of motor neurons have yet to be elucidated. Here, retinoic acid and sonic hedgehog were applied to examine whether hADSCs could be differentiated into motor neurons. As early as 6 h after induction, hADSCs were changed toward neuronal morphology. After induction, hADSCs showed positive immunocytochemical staining for β-III-tubulin, choline acetyltransferase, and neuron-specific enolase. Reverse-transcriptase polymerase chain reaction characterization indicated that cells differentiated from hADSCs were restricted to the ventral spinal fate (Nkx2.2, Pax6, Hb9, and Olig2). Our results suggest that hADSCs may be a potential candidate in cellular therapy for motor neuron disease.

PMID: 21532392 [PubMed - indexed for MEDLINE]


A PEGylated fibrin-based wound dressing with antimicrobial and angiogenic activity.
Source:  Pub Med
Monday, 29 November 1999 17:00

A PEGylated fibrin-based wound dressing with antimicrobial and angiogenic activity.

Acta Biomater. 2011 Jul;7(7):2787-96

Authors: Seetharaman S, Natesan S, Stowers RS, Mullens C, Baer DG, Suggs LJ, Christy RJ

Abstract
Wounds sustained under battlefield conditions are considered to be contaminated and their initial treatment should focus on decreasing this contamination and thus reducing the possibility of infection. The early and aggressive administration of antimicrobial treatment starting with intervention on the battlefield has resulted in improved patient outcomes and is considered the standard of care. Chitosan microspheres (CSM) loaded with silver sulfadiazine (SSD) were developed via a novel water-in-oil emulsion technique to address this problem. The SSD-loaded spheres were porous with needle-like structures (attributed to SSD) that were evenly distributed over the spheres. The average particle size of the SSD-CSM was 125-180 μm with 76.50 ± 2.8% drug entrapment. As a potential new wound dressing with angiogenic activity SSD-CSM particles were impregnated in polyethylene glycol (PEGylated) fibrin gels. In vitro drug release studies showed that a burst release of 27.02% in 6h was achieved, with controlled release for 72 h, with an equilibrium concentration of 27.7% (70 μg). SSD-CSM-PEGylated fibrin gels were able to exhibit microbicidal activity at 125 and 100 μg ml(-1) against Staphylococcus aureus and Pseudomonas aeruginosa, respectively. The in vitro vasculogenic activity of this composite dressing was shown by seeding adipose-derived stem cells (ASC) in SSD-CSM-PEGylated fibrin gels. The ASC spontaneously formed microvascular tube-like structures without the addition of any exogenous factors. This provides a method for the extended release of an antimicrobial drug in a matrix that may provide an excellent cellular environment for revascularization of infected wounds.

PMID: 21515420 [PubMed - indexed for MEDLINE]


Adipose-derived mesenchymal stem cell protects kidneys against ischemia-reperfusion injury through suppressing oxidative stress and inflammatory reaction.
Source:  Pub Med
Monday, 29 November 1999 17:00

Adipose-derived mesenchymal stem cell protects kidneys against ischemia-reperfusion injury through suppressing oxidative stress and inflammatory reaction.

J Transl Med. 2011;9:51

Authors: Chen YT, Sun CK, Lin YC, Chang LT, Chen YL, Tsai TH, Chung SY, Chua S, Kao YH, Yen CH, Shao PL, Chang KC, Leu S, Yip HK

Abstract
BACKGROUND: Reactive oxygen species are important mediators exerting toxic effects on various organs during ischemia-reperfusion (IR) injury. We hypothesized that adipose-derived mesenchymal stem cells (ADMSCs) protect the kidney against oxidative stress and inflammatory stimuli in rat during renal IR injury.
METHODS: Adult male Sprague-Dawley (SD) rats (n = 24) were equally randomized into group 1 (sham control), group 2 (IR plus culture medium only), and group 3 (IR plus immediate intra-renal administration of 1.0 × 106 autologous ADMSCs, followed by intravenous ADMSCs at 6 h and 24 h after IR). The duration of ischemia was 1 h, followed by 72 hours of reperfusion before the animals were sacrificed.
RESULTS: Serum creatinine and blood urea nitrogen levels and the degree of histological abnormalities were markedly lower in group 3 than in group 2 (all p < 0.03). The mRNA expressions of inflammatory, oxidative stress, and apoptotic biomarkers were lower, whereas the anti-inflammatory, anti-oxidative, and anti-apoptotic biomarkers were higher in group 3 than in group 2 (all p < 0.03). Immunofluorescent staining showed a higher number of CD31+, von Willebrand Factor+, and heme oxygenase (HO)-1+ cells in group 3 than in group 2 (all p < 0.05). Western blot showed notably higher NAD(P)H quinone oxidoreductase 1 and HO-1 activities, two indicators of anti-oxidative capacity, in group 3 than those in group 2 (all p < 0.04). Immunohistochemical staining showed higher glutathione peroxidase and glutathione reductase activities in group 3 than in group 2 (all p < 0.02)
CONCLUSION: ADMSC therapy minimized kidney damage after IR injury through suppressing oxidative stress and inflammatory response.

PMID: 21545725 [PubMed - indexed for MEDLINE]


Proliferation and differentiation potential of human adipose-derived mesenchymal stem cells isolated from elderly patients with osteoporotic fractures.
Source:  Pub Med
Monday, 29 November 1999 17:00

Proliferation and differentiation potential of human adipose-derived mesenchymal stem cells isolated from elderly patients with osteoporotic fractures.

J Cell Mol Med. 2011 May 5;

Authors: Chen HT, Lee MJ, Chen CH, Chuang SC, Chang LF, Ho ML, Hung SH, Fu YC, Wang YH, Wang HI, Wang GJ, Kang L, Chang JK

Abstract
Aging has less effect on adipose-derived mesenchymal stem cells (ADSCs) than on bone marrow-derived mesenchymal stem cells (BMSCs), but whether the fact holds true in stem cells from elderly patients with osteoporotic fractures is unknown. In this study, ADSCs and BMSCs of the same donor were harvested and divided into two age groups. Group A consisted of 14 young patients (36.4 ± 11.8 years old), and group B consisted of 8 elderly patients (71.4 ± 3.6 years old) with osteoporotic fractures. We found that the doubling time of ADSCs from both age groups was maintained below 70 hrs, while that of BMSCs increased significantly with the number of passage. When ADSCs and BMSCs from the same patient was compared, there was a significant increase in the doubling time of BMSCs in each individual from passages 3 to 6. Upon osteogenic induction, the level of matrix mineralization of ADSCs from group B was comparable to that of ADSCs from group A, while BMSCs from group B produced least amount of mineral deposits and had a lower expression level of osteogenic genes. The p21 gene expression and senescence-associated β-galactosidase activity were lower in ADSCs compared to BMSCs, which may be partly responsible for the greater proliferation and differentiation potential of ADSCs. It is concluded that the proliferation and osteogenic differentiation of ADSCs were less affected by age and multiple passage than BMSCs, suggesting that ADSCs may become a potentially effective therapeutic option for cell-based therapy, especially in elderly patients with osteoporosis.

PMID: 21545685 [PubMed - as supplied by publisher]


[Comparison of chondrogenic differentiation of adipose tissue-derived mesenchymal stem cells with cultured chondrocytes and bone marrow mesenchymal stem cells].
Source:  Pub Med
Monday, 29 November 1999 17:00

[Comparison of chondrogenic differentiation of adipose tissue-derived mesenchymal stem cells with cultured chondrocytes and bone marrow mesenchymal stem cells].

Acta Chir Orthop Traumatol Cech. 2011;78(2):138-44

Authors: Havlas V, Kos P, Jendelová P, Lesný P, Trč T, Syková E

Abstract
PURPOSE OF THE STUDY: Many congenital and acquired disorders as well as sequelae of injury are associated with articular cartilage degeneration, which adversely affects the patient's quality of life. The currently used cell therapy with cultured chondrocytes has its disadvantages due to a process of de-differentiation of chondrocytes during cultivation. We believe that the mesenchymal stem cell therapy offers a new treatment options.
MATERIAL AND METHODS: The adult mesenchymal stem cells (MSCs) for chondrocyte differentiation are usually obtained from bone marrow mesenchymal stem cells (BMSCs). In this study these cells were compared with mesenchymal stem cells derived from adipose tissue (AMSCs). The aim of the study was to verify the ability of human BMSCs and AMSCs to differentiate into chondrocytes in vitro in the presence or absence of transforming growth factor beta (TGF-ß1). Human BMSCs and AMSCs were collected from healthy donors during orthopaedic surgeries, in vitro cultured in three passages to obtain the required quantity of cells. A pellet culture system was used for chondrocyte differentiation.
RESULTS: At 21 days of cultivation, cell aggregates grown in the chondrogenic medium were larger than those cultured in the control medium. Both the BMSCs and AMSCs pellet cultures showed spontaneous chondrogenesis. Histological staining with haematoxylin and eosin and Masson's trichrome stains, as well as immunohistochemical staining to detect type II collagen revealed no apparent differences between the pellet cultures with TGF-ß1 presence and those without it. The real-time RT-PCR detected expression of the type II collagen gene in all tested cultures. In the BMSCs pellet culture only, TGF-ß1 presence resulted in a decrease in type I collagen mRNA levels and in an increase in type II collagen mRNA values.
DISCUSSION: Our results showed an in vitro chondrogenic potential of mature human mesenchymal stem cells derived from both bone marrow and adipose tissue. In agreement with the relevant literature data, we suggest that both cell types have an equal prospect for use in cartilage tissue engineering.

PMID: 21575557 [PubMed - indexed for MEDLINE]


Transduction of human adipose-derived mesenchymal stem cells by recombinant adeno-associated virus vectors.
Source:  Pub Med
Monday, 29 November 1999 17:00

Transduction of human adipose-derived mesenchymal stem cells by recombinant adeno-associated virus vectors.

Tissue Eng Part C Methods. 2011 Sep;17(9):949-59

Authors: Locke M, Ussher JE, Mistry R, Taylor JA, Dunbar PR

Abstract
Human adipose-derived stem cells (ASCs) are attractive targets for genetic manipulation and cellular therapies. However, current methods of gene transfer are limited by lack of efficiency, toxicity, or safety concerns. Recombinant adeno-associated virus (rAAV) has been extensively assessed as a gene therapy vector and has an excellent safety profile. This study reports the efficient transduction of well-characterized, homogeneous cultures of human ASCs by rAAV serotypes 2, 5, and 6. Transduction with rAAV2 at high multiplicity of infection was associated with reduced cell viability; however, no adverse effect was seen with serotypes 5 and 6. A further increase in transduction efficiency was observed using a rAAV6 Y731F tyrosine capsid mutant. rAAV-transduced ASCs retained their adipogenic potential. Therefore, rAAV serotypes 2, 5, and 6 should be considered the vectors of choice for genetic manipulation of ASCs.

PMID: 21563982 [PubMed - in process]


Stearidonic and eicosapentaenoic acids inhibit interleukin-6 expression in ob/ob mouse adipose stem cells via Toll-like receptor-2-mediated pathways.
Source:  Pub Med
Monday, 29 November 1999 17:00

Stearidonic and eicosapentaenoic acids inhibit interleukin-6 expression in ob/ob mouse adipose stem cells via Toll-like receptor-2-mediated pathways.

J Nutr. 2011 Jul;141(7):1260-6

Authors: Hsueh HW, Zhou Z, Whelan J, Allen KG, Moustaid-Moussa N, Kim H, Claycombe KJ

Abstract
Increased adipose tissue positively correlates with circulating inflammatory cytokines such as IL-6. We previously reported that adipose stem cells from genetically obese ob/ob mice produce significantly higher levels of IL-6 compared with other cell types such as adipocytes and macrophages within adipose tissue. We also demonstrated that (n-3) PUFA have antiinflammatory effects on adipocyte IL-6 secretion. Based on these findings, we hypothesized that EPA [20:5 (n-3)] and stearidonic acid [SDA, 18:4 (n-3)] would decrease LPS (200 μg/L)-induced IL-6 secretion and IL-6 mRNA content in the adipose stem cells. SDA (100 μmol/L) and EPA (100 μmol/L) significantly reduced LPS-induced IL-6 secretion and decreased IL-6 mRNA expression. To determine the underlying intracellular mechanisms, we tested whether LPS-induced Toll-like-receptor (TLR) 4 and TLR2 expression were modulated by these fatty acids using Western-blot analysis. EPA and SDA suppressed LPS-induced TLR2 but not TLR4 protein expression in the adipose stem cells. Furthermore, SDA and EPA significantly lowered the activation and translocation of NF-κB, a TLR2 downstream signaling target, while protein expression of extracellular signal-regulated kinases-1/2 were unaffected. Collectively, our results suggest that EPA and SDA inhibit LPS-induced IL-6 secretion and IL-6 mRNA expression in the adipose stem cells by decreasing TRL2-mediated signaling pathways.

PMID: 21562237 [PubMed - indexed for MEDLINE]


Differential requirement for utrophin in the induced pluripotent stem cell correction of muscle versus fat in muscular dystrophy mice.
Source:  Pub Med
Monday, 29 November 1999 17:00

Differential requirement for utrophin in the induced pluripotent stem cell correction of muscle versus fat in muscular dystrophy mice.

PLoS One. 2011;6(5):e20065

Authors: Beck AJ, Vitale JM, Zhao Q, Schneider JS, Chang C, Altaf A, Michaels J, Bhaumik M, Grange R, Fraidenraich D

Abstract
Duchenne muscular dystrophy (DMD) is an incurable degenerative muscle disorder. We injected WT mouse induced pluripotent stem cells (iPSCs) into mdx and mdx∶utrophin mutant blastocysts, which are predisposed to develop DMD with an increasing degree of severity (mdx <<< mdx∶utrophin). In mdx chimeras, iPSC-dystrophin was supplied to the muscle sarcolemma to effect corrections at morphological and functional levels. Dystrobrevin was observed in dystrophin-positive and, at a lesser extent, utrophin-positive areas. In the mdx∶utrophin mutant chimeras, although iPSC-dystrophin was also supplied to the muscle sarcolemma, mice still displayed poor skeletal muscle histopathology, and negligible levels of dystrobrevin in dystrophin- and utrophin-negative areas. Not only dystrophin-expressing tissues are affected by iPSCs. Mdx and mdx∶utrophin mice have reduced fat/body weight ratio, but iPSC injection normalized this parameter in both mdx and mdx∶utrophin chimeras, despite the fact that utrophin was compromised in the mdx∶utrophin chimeric fat. The results suggest that the presence of utrophin is required for the iPSC-corrections in skeletal muscle. Furthermore, the results highlight a potential (utrophin-independent) non-cell autonomous role for iPSC-dystrophin in the corrections of non-muscle tissue like fat, which is intimately related to the muscle.

PMID: 21603573 [PubMed - indexed for MEDLINE]


"Humanized" stem cell culture techniques: the animal serum controversy.
Source:  Pub Med
Monday, 29 November 1999 17:00

"Humanized" stem cell culture techniques: the animal serum controversy.

Stem Cells Int. 2011;2011:504723

Authors: Tekkatte C, Gunasingh GP, Cherian KM, Sankaranarayanan K

Abstract
Cellular therapy is reaching a pinnacle with an understanding of the potential of human mesenchymal stem cells (hMSCs) to regenerate damaged tissue in the body. The limited numbers of these hMSCs in currently identified sources, like bone marrow, adipose tissue, and so forth, bring forth the need for their in vitro culture/expansion. However, the extensive usage of supplements containing xenogeneic components in the expansion-media might pose a risk to the post-transplantation safety of patients. This warrants the necessity to identify and develop chemically defined or "humanized" supplements which would make in vitro cultured/processed cells relatively safer for transplantation in regenerative medicine. In this paper, we outline the various caveats associated with conventionally used supplements of xenogenic origin and also portray the possible alternatives/additives which could one day herald the dawn of a new era in the translation of in vitro cultured cells to therapeutic interventions.

PMID: 21603148 [PubMed]


Inhibition of osteogenic differentiation of human adipose-derived stromal cells by retinoblastoma binding protein 2 repression of RUNX2-activated transcription.
Source:  Pub Med
Monday, 29 November 1999 17:00

Inhibition of osteogenic differentiation of human adipose-derived stromal cells by retinoblastoma binding protein 2 repression of RUNX2-activated transcription.

Stem Cells. 2011 Jul;29(7):1112-25

Authors: Ge W, Shi L, Zhou Y, Liu Y, Ma GE, Jiang Y, Xu Y, Zhang X, Feng H

Abstract
Histone methylation is regarded as an important type of histone modification defining the epigenetic program during the lineage differentiation of stem cells. A better understanding of this epigenetic mechanism that governs osteogenic differentiation of human adipose-derived stromal cells (hASCs) can improve bone tissue engineering and provide new insights into the modulation of hASC-based cell therapy. Retinoblastoma binding protein 2 (RBP2) is a histone demethylase that specifically catalyzes demethylation of dimethyl or trimethyl histone H3 lysine 4 (H3K4me2 or H3K4me3), which is normally associated with transcriptionally active genes. In this study, the roles of RBP2 in osteogenic differentiation of hASCs were investigated. We found that RBP2 knockdown by lentiviruses expressing small interfering RNA promoted osteogenic differentiation of hASCs in vitro and in vivo. In addition, we demonstrated that knockdown of RBP2 resulted in marked increases of mRNA expression of osteogenesis-associated genes such as alkaline phosphatase (ALP), osteocalcin (OC), and osterix (OSX). RBP2 was shown to occupy the promoters of OSX and OC to maintain the level of the H3K4me3 mark by chromatin immunoprecipitation assays. Furthermore, coimmunoprecipitation and luciferase reporter experiments suggested that RBP2 was physically and functionally associated with RUNX2, an essential transcription factor that governed osteoblastic differentiation. Significantly, RUNX2 knockdown impaired the repressive activity of RBP2 in osteogenic differentiation of hASCs. Altogether, our study is the first to demonstrate the functional and biological roles of H3K4 demethylase RBP2 in osteogenic differentiation of hASCs and to link RBP2 to the transcriptional regulation of RUNX2.

PMID: 21604327 [PubMed - in process]


Editorial comment.
Source:  Pub Med
Monday, 29 November 1999 17:00

Editorial comment.

Urology. 2011 Jun;77(6):e3; author reply e4

Authors: Frauscher F, Marksteiner R

PMID: 21624585 [PubMed - indexed for MEDLINE]


[Experimental study on adipose-derived stem cells combined with acellular dermal matrix particles for vocal fold injection].
Source:  Pub Med
Monday, 29 November 1999 17:00

[Experimental study on adipose-derived stem cells combined with acellular dermal matrix particles for vocal fold injection].

Zhonghua Er Bi Yan Hou Tou Jing Wai Ke Za Zhi. 2011 Apr;46(4):283-7

Authors: Liang Q, Liu Y, Deng ZH, Li XB, Li XF, Zhao Y, Liang YY, Jin Y

Abstract
OBJECTIVE: To explore the feasibility of adipose-derived stem cells (ASC) combined with micronized acellular dermal matrix (MADM) for vocal cord injection.
METHODS: The adipose-deprived stem cells were harvested from rabbit adipose tissue in vitro. The 3rd generation of ASC was labeled with DiI (1,1-dioctadecyl-3,3,3,3-tetramethylindocarbocyanine perchlorate) and cultured with MADM to form a complex. The adhesion of ASC to MADM was observed by fluorescence microscope and electron microscope. The proliferation of ASC on MADM was evaluated by 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxy methoxyphenyl)-2-(4-sulfonyl)-2H-tetrazolium, inner salt (MTS). Three days after the culture, the complex was mixed with appropriate amount of collagen, and then injected into the unilateral vocal cord of the rabbit. The animals were sacrificed 2, 4, 8 weeks after injection, the survival time and distribution of ASC in vocal fold were tested, and the responses of vocal cord to ASC-MADM and the degradation of MADM were observed.
RESULTS: The ASC adhered to MADM and grew well (P < 0.05 or < 0.01), showing good compatibility with MADM in vocal cord tissue. The complex of ASC-MADM could be injected into the rabbit vocal cords, while no adverse reactions was observed in the vocal cord by endoscope, frozen section and HE staining. ASC could survive for 8 weeks in vocal cords, and no inflammatory cell infiltration was observed.
CONCLUSIONS: MADM is an ideal scaffold material and shows perfect compatibility with ASC which can adhere and proliferate well on it. The complex of ASC-MADM can be injected into the vocal cord and can survive. There is no adverse reaction in vocal cords.

PMID: 21624246 [PubMed - indexed for MEDLINE]


Stem cell induced cardiac regeneration: fusion/mitochondrial exchange and/or transdifferentiation?
Source:  Pub Med
Monday, 29 November 1999 17:00

Stem cell induced cardiac regeneration: fusion/mitochondrial exchange and/or transdifferentiation?

Cell Cycle. 2011 Jul 15;10(14):2281-6

Authors: Song YH, Pinkernell K, Alt E

Abstract
Potentially, adult stem cell-based therapy provides a new therapeutic option for myocardial regeneration. However, to date, with regard to the benefits seen, the mechanisms involved in stem cell-based therapy are not well understood. Suggested pathways proposed so far include fusion of stem cells with cardiomyocytes, transdifferentiation into cardiac and vascular cells and secretion of paracrine factors. In a recent study, our group examined the fate of human adipose tissue-derived stem cells (hASCs) fused with rat cardiomyocytes after treatment with fusion-inducing hemagglutinating virus of Japan (HVJ). In this study, we demonstrated that cells of fused hASC cardiomyocytes display a cardiomyocyte phenotype and spontaneous rhythmic contraction and generate an action potential in vitro. As part of the work underlying this paper, we co-cultured rat neonatal cardiomyocytes with hASCs or pig bone marrow-derived mesenchymal stem cells (MSCs), where ASCs or MSCs had previously been transduced with a lentivirus encoding eGFP. Our data evidence early cardiac contractile proteins, such as Titin and MF20, identified in eGFP-positive cells, suggesting a cardiomyogenic phenotype. Recent work by others has shown that the myogenic conversion increased when BMSCs were cultured with apoptotic cells. In this Extra View article, we review the current understanding of stem cell-derived factors, fusion/partial fusion and the manner in which the exchange of cellular contents between stem cells and cardiomyocytes might contribute to the reprogramming of fully differentiated cardiomyocytes based on recently published literature.

PMID: 21654195 [PubMed - in process]


Efficacy of Adipose Tissue-Derived Mesenchymal Stem Cells for Fulminant Hepatitis in Mice Induced by Concanavalin A.
Source:  Pub Med
Monday, 29 November 1999 17:00

Efficacy of Adipose Tissue-Derived Mesenchymal Stem Cells for Fulminant Hepatitis in Mice Induced by Concanavalin A.

J Gastroenterol Hepatol. 2011 Jun 7;

Authors: Kubo N, Narumi S, Kijima H, Mizukami H, Yagihashi S, Hakamada K, Nakane A

Abstract
Background and Aim: Fulminant hepatitis is mainly caused by excessive immune response-mediated liver injury and its definitive therapy is liver transplantation. Mesenchymal stem cells, one of the adult stem cells, have an immunomodulatory effect on immune cells and reside in various tissues. The aim of this study was to investigate a therapeutic effect of adipose tissue-derived mesenchymal stem cells (ASCs) on fulminant hepatitis induced by concanavalin A (ConA). Methods: ASCs were isolated from adipose tissues of BALB/c mice and confirmed by detection of cell surface markers and induction of multi-lineage differentiation. BALB/c mice were injected with ConA and treated with ASCs, phosphate buffered saline (PBS) or splenocytes (SPLCs). Survival rates, levels of serum liver enzymes, titers of serum cytokines, histopathology and localization of ASCs were investigated. Result: The survival rate of ASC-injected mice significantly increased compared to PBS or SPLC-injected mice. This effect was dependent on doses and timing of ASCs injected. Improvement of liver enzyme levels, histopathological changes and suppression of inflammatory cytokine production were observed in ASC-injected mice. Fluorescent stained ASCs were detected in inflammatory liver, but not in normal liver. Conclusion: These results suggest that ASC treatment has a high potential to be an innovative therapy for fulminant hepatitis.

PMID: 21649723 [PubMed - as supplied by publisher]


Chitosan/silk fibroin-based tissue-engineered graft seeded with adipose-derived stem cells enhances nerve regeneration in a rat model.
Source:  Pub Med
Monday, 29 November 1999 17:00

Chitosan/silk fibroin-based tissue-engineered graft seeded with adipose-derived stem cells enhances nerve regeneration in a rat model.

J Mater Sci Mater Med. 2011 Aug;22(8):1947-64

Authors: Wei Y, Gong K, Zheng Z, Wang A, Ao Q, Gong Y, Zhang X

Abstract
Sciatic nerve injury presents an ongoing challenge in reconstructive surgery. Local stem cell application has recently been suggested as a possible novel therapy. In the present study we evaluated the potential of a chitosan/silk fibroin scaffold serving as a delivery vehicle for adipose-derived stem cells and as a structural framework for the injured nerve regeneration. The cell-loaded scaffolds were used to regenerate rat sciatic nerve across a 10 mm surgically-induced sciatic nerve injury. The functional nerve recovery was assessed by both walking track and histology analysis. Results showed that the reconstruction of the injured sciatic nerve had been significantly enhanced with restoration of nerve continuity and function recovery in the cell-loaded scaffold groups, and their target skeletal muscle had been extensively reinnervated. This study raises a potential possibility of using the newly developed nerve grafts as a promising alternative for nerve regeneration.

PMID: 21656031 [PubMed - in process]


Competitive electroporation formulation for cell therapy.
Source:  Pub Med
Monday, 29 November 1999 17:00

Competitive electroporation formulation for cell therapy.

Cancer Gene Ther. 2011 Aug;18(8):579-86

Authors: Flanagan M, Gimble JM, Yu G, Wu X, Xia X, Hu J, Yao S, Li S

Abstract
Established cell transfection via nucleofection relies on nucleofection buffers with unknown and proprietary makeup due to trade secrecy, inhibiting the possibility of using this otherwise effective method for developing cell therapy. We devised a three-step method for discovering an optimal formulation for the nucleofection of any cell line. These steps include the selection of the best nucleofection program and known buffer type, selection of the best polymer for boosting the transfection efficiency of the best buffer and the comparison with the optimal buffer from an established commercial vendor (Amaxa). Using this three-step selection system, competitive nucleofection formulations were discovered for multiple cell lines, which are equal to or surpass the efficiency of the Amaxa nucleofector solution in a variety of cells and cell lines, including primary adipose stem cells, muscle cells, tumor cells and immune cells. Through the use of scanning electron microscopy, we have revealed morphological changes, which predispose for the ability of these buffers to assist in transferring plasmid DNA into the nuclear space. Our formulation may greatly reduce the cost of electroporation study in laboratory and boosts the potential of application of electroporation-based cell therapies in clinical trials.

PMID: 21660061 [PubMed - in process]


Combined gene and stem cell therapy for cutaneous wound healing.
Source:  Pub Med
Monday, 29 November 1999 17:00

Combined gene and stem cell therapy for cutaneous wound healing.

Mol Pharm. 2011 Oct 3;8(5):1471-9

Authors: Gauglitz GG, Jeschke MG

Abstract
In current medical practice, wound therapy remains a clinical challenge and much effort has been focused on the development of novel therapeutic approaches for wound treatment. Gene therapy, initially developed for treatment of congenital defects, represents a promising option for enhancing wound repair. In order to accelerate wound closure, genes encoding for growth factors or cytokines have shown the most potential. The majority of gene delivery systems are based on viral transfection, naked DNA application, high pressure injection, and liposomal vectors. Besides advances stemming from breakthroughs in recombinant growth factors and bioengineered skin, there has been a significant increase in the understanding of stem cell biology in the field of cutaneous wound healing. A variety of sources, such as bone marrow, umbilical cord blood, adipose tissue and skin/hair follicles, have been utilized to isolate stem cells and to modulate the healing response of acute and chronic wounds. Recent data have demonstrated the feasibility of autologous adult stem cell therapy in cutaneous repair and regeneration. Very recently, stem cell based skin engineering in conjunction with gene recombination, in which the stem cells act as both the seed cells and the vehicle for gene delivery to the wound site, represents the most attractive field for generating a regenerative strategy for wound therapy. The aim of this article is to discuss the use and the potential of these novel technologies in order to improve wound healing capacities.

PMID: 21657247 [PubMed - in process]


Stem cells and growth factor delivery systems for cardiovascular disease.
Source:  Pub Med
Monday, 29 November 1999 17:00

Stem cells and growth factor delivery systems for cardiovascular disease.

J Biotechnol. 2011 Jul 20;154(4):291-7

Authors: Madonna R, De Caterina R

Abstract
Coronary (CAD) and peripheral (PAD) artery diseases are major causes of morbidity and mortality, and millions of CAD and PAD patients are treated by various medications, bypass surgery or angioplasty around the world. Such patients might benefit from novel stem cells and tissue engineering strategies aimed at accelerating natural processes of postnatal collateral vessel formation and repairing damaged tissues. By combining three fundamental "tools", namely stem cells, biomaterials and growth factors (GFs), such strategies may enhance the efficacy of cell therapy in several ways: (a) by supplying exogenous stem cells or GFs that stimulate resident cardiac stem cell (CSC) migration, engraftment and commitment to cardiomyocytes, and that induce and modulate arterial response to ischemia; (b) by supporting the maintenance of GFs and transplanted stem cells in the damaged tissues through the use of biocompatible and biodegradable polymers for a period of time sufficient to allow histological and anatomical restoration of the damaged tissue. This review will discuss the potential of combining stem cells and new delivery systems for growth factors, such as vehicle-based delivery strategies or cell-based gene therapy, to facilitate regeneration of ischemic tissues. These approaches would promote the ability of resident CSCs or of exogenous multipotent stem cells such as adipose tissue-derived mesenchymal stem cells (AT-MSCs) to induce the healing of damaged tissue, by recruiting and directing these cells into the damage area and by improving angiogenesis and reperfusion of ischemic tissues.

PMID: 21663773 [PubMed - in process]


Pigment Epithelial-Derived Factor and Melanoma Differentiation Associated Gene-7 Cytokine Gene Therapies Delivered by Adipose-Derived Stromal/Mesenchymal Stem Cells are Effective in Reducing Prostate Cancer Cell Growth.
Source:  Pub Med
Monday, 29 November 1999 17:00

Pigment Epithelial-Derived Factor and Melanoma Differentiation Associated Gene-7 Cytokine Gene Therapies Delivered by Adipose-Derived Stromal/Mesenchymal Stem Cells are Effective in Reducing Prostate Cancer Cell Growth.

Stem Cells Dev. 2011 Jul 26;

Authors: Zolochevska O, Yu G, Gimble JM, Figueiredo ML

Abstract
Adipose-derived stromal/mesenchymal stem cells (ASC) have gained interest as promising tools for delivering cancer therapy. Adipose tissue can be obtained readily in amounts sufficient for ASC isolation, which can be expanded rapidly, allowing its use at low passage numbers, and can be transduced by viral and nonviral means. Our goal was to examine the potential of ASC to deliver cytokine gene therapies melanoma differentiation associated gene-7 (MDA-7) or pigment epithelial-derived factor (PEDF) to cancer cells. These novel cytokines are a potent proapoptotic and an antiangiogenesis mediator, respectively, with potential as antitumor agents. Expression of cytokine therapies did not adversely affect ASC biology, and these cells were still able to differentiate and retain normal viability. The ASC cytokine therapies were efficient in reducing tumor cell growth in coculture and also in suppressing in vitro angiogenesis phenotypes. We also observed that ASC retained their innate ability to migrate toward tumor cells in coculture, and this ability could be blocked by inhibition of CXCR4 signaling. The ASC were found to be nontumorigenic in vitro using a soft agar assay, as well as in vivo, utilizing 2 prostate cancer xenograft models. The ASC-MDA7 only reduced tumor growth in the TRAMP-C2-Ras (TC2Ras) prostate cancer model. The ASC-PEDF, however, reduced growth in both the TC2Ras and the PC3 highly aggressive prostate cancer models, and it was able to completely prevent prostate tumor establishment in vivo. In conclusion, ASC expressing PEDF and MDA7 could effectively reduce prostate tumor growth in vivo, suggesting ASC-cytokine therapies might have translational applications, especially the PEDF modality.

PMID: 21671747 [PubMed - as supplied by publisher]


Human adipose tissue-derived mesenchymal stem cells: Characteristics and therapeutic potential as cellular vehicles for prodrug gene therapy against brainstem gliomas.
Source:  Pub Med
Monday, 29 November 1999 17:00

Human adipose tissue-derived mesenchymal stem cells: Characteristics and therapeutic potential as cellular vehicles for prodrug gene therapy against brainstem gliomas.

Eur J Cancer. 2011 Jun 8;

Authors: Choi SA, Lee JY, Wang KC, Phi JH, Song SH, Song J, Kim SK

Abstract
Human mesenchymal stem cells (hMSCs) have emerged as attractive cellular vehicles for gene therapy against brain malignancy because of their targeted tropism for cancer and the intrinsic attribute of autologous transplantation. We evaluated the characteristics and therapeutic potential of human adipose tissue-derived MSCs (hAT-MSCs) and prodrug gene therapy against diffuse pontine gliomas. The hAT-MSCs were isolated from human adipose tissue and characterised for morphology, surface markers and potential to differentiate into mesenchymal and neuronal lineages. We genetically modified hAT-MSCs to express rabbit carboxylesterase (rCE) enzyme, which can efficiently convert the prodrug CPT-11 (irinotecan-7-ethyl-10-[4-(1-piperidino)-1-piperidino]carbonyloxycamptothecin), into the active drug SN-38 (7-ethyl-10-hydroxycamptothecin). The migratory capacity of hAT-MSCs expressing rCE (hAT-MSC.rCE), their ability to convert CPT-11 to SN-38 and cytotoxic effect on F98 cells were evaluated in vitro. The therapeutic potential of hAT-MSC.rCE was confirmed using a rat brainstem glioma model. The hAT-MSCs showed fibroblast-like morphology and expressed hMSC-specific markers including CD73, CD90 and CD105. The hAT-MSCs could differentiate into a mesenchymal lineage and transdifferentiate into a neuronal lineage under optimum culture conditions. The hAT-MSC.rCE converted CPT-11 to SN-38 and preserved the tumour tropism of hAT-MSCs. Brainstem glioma-bearing rats treated with hAT-MSC.rCE and CPT-11 survived 5d more than rats treated with CPT-11 only (p=0.0018). Our study demonstrates that hAT-MSCs can be easily prepared and genetically modified as cellular vehicles for prodrug gene therapy and that they have therapeutic potential against brainstem gliomas.

PMID: 21664124 [PubMed - as supplied by publisher]


Islet-like cell aggregates generated from human adipose tissue derived stem cells ameliorate experimental diabetes in mice.
Source:  Pub Med
Monday, 29 November 1999 17:00

Islet-like cell aggregates generated from human adipose tissue derived stem cells ameliorate experimental diabetes in mice.

PLoS One. 2011;6(6):e20615

Authors: Chandra V, Swetha G, Muthyala S, Jaiswal AK, Bellare JR, Nair PD, Bhonde RR

Abstract
BACKGROUND: Type 1 Diabetes Mellitus is caused by auto immune destruction of insulin producing beta cells in the pancreas. Currently available treatments include transplantation of isolated islets from donor pancreas to the patient. However, this method is limited by inadequate means of immuno-suppression to prevent islet rejection and importantly, limited supply of islets for transplantation. Autologous adult stem cells are now considered for cell replacement therapy in diabetes as it has the potential to generate neo-islets which are genetically part of the treated individual. Adopting methods of islet encapsulation in immuno-isolatory devices would eliminate the need for immuno-suppressants.
METHODOLOGY/PRINCIPAL FINDINGS: In the present study we explore the potential of human adipose tissue derived adult stem cells (h-ASCs) to differentiate into functional islet like cell aggregates (ICAs). Our stage specific differentiation protocol permit the conversion of mesodermic h-ASCs to definitive endoderm (Hnf3β, TCF2 and Sox17) and to PDX1, Ngn3, NeuroD, Pax4 positive pancreatic endoderm which further matures in vitro to secrete insulin. These ICAs are shown to produce human C-peptide in a glucose dependent manner exhibiting in-vitro functionality. Transplantation of mature ICAs, packed in immuno-isolatory biocompatible capsules to STZ induced diabetic mice restored near normoglycemia within 3-4 weeks. The detection of human C-peptide, 1155±165 pM in blood serum of experimental mice demonstrate the efficacy of our differentiation approach.
CONCLUSIONS: h-ASC is an ideal population of personal stem cells for cell replacement therapy, given that they are abundant, easily available and autologous in origin. Our findings present evidence that h-ASCs could be induced to differentiate into physiologically competent functional islet like cell aggregates, which may provide as a source of alternative islets for cell replacement therapy in type 1 diabetes.

PMID: 21687731 [PubMed - indexed for MEDLINE]


Incorporating platelet-rich plasma into electrospun scaffolds for tissue engineering applications.
Source:  Pub Med
Monday, 29 November 1999 17:00

Incorporating platelet-rich plasma into electrospun scaffolds for tissue engineering applications.

Tissue Eng Part A. 2011 Nov;17(21-22):2723-37

Authors: Sell SA, Wolfe PS, Ericksen JJ, Simpson DG, Bowlin GL

Abstract
Platelet-rich plasma (PRP) therapy has seen a recent spike in clinical interest due to the potential that the highly concentrated platelet solutions hold for stimulating tissue repair and regeneration. The aim of this study was to incorporate PRP into a number of electrospun materials to determine how growth factors are eluted from the structures, and what effect the presence of these factors has on enhancing electrospun scaffold bioactivity. PRP underwent a freeze-thaw-freeze process to lyse platelets, followed by lyophilization to create a powdered preparation rich in growth factors (PRGF), which was subsequently added to the electrospinning process. Release of protein from scaffolds over time was quantified, along with the quantification of human macrophage and adipose-derived stem cell (ADSC) chemotaxis and proliferation. Protein assays demonstrated a sustained release of protein from PRGF-containing scaffolds at up to 35 days in culture. Scaffold bioactivity was enhanced as ADSCs demonstrated increased proliferation in the presence of PRGF, whereas macrophages demonstrated increased chemotaxis to PRGF. In conclusion, the work performed in this study demonstrated that the incorporation of PRGF into electrospun structures has a significant positive influence on the bioactivity of the scaffolds, and may prove beneficial in a number of tissue engineering applications.

PMID: 21679135 [PubMed - in process]


The source of human mesenchymal stromal cells influences their TLR profile as well as their functional properties.
Source:  Pub Med
Monday, 29 November 1999 17:00

The source of human mesenchymal stromal cells influences their TLR profile as well as their functional properties.

Cell Immunol. 2011;270(2):207-16

Authors: Raicevic G, Najar M, Stamatopoulos B, De Bruyn C, Meuleman N, Bron D, Toungouz M, Lagneaux L

Abstract
Mesenchymal stromal cells (MSC) can be expanded from different sources. We compared the influence of inflammation and TLR ligation on the phenotype and function of MSC derived from bone marrow (BM), adipose tissue (AT), and Wharton's jelly (WJ). WJ-MSC were featured by a lack of TLR4 expression. While inflammation upregulated TLR3 in all three MSC types, TLR4 upregulation was observed only on BM-MSC. TLR ligation increased the production of inflammatory cytokines in BM- and AT-MSC but not in WJ-MSC and augmented anti-inflammatory cytokines in AT-MSC. Although inflammation increased in all MSC types the secretion of inflammatory cytokines, additional TLR triggering did not have further effect on WJ-MSC. The immunosuppressive potential of WJ-MSC on MLR was affected neither by inflammation nor by TLR triggering. This resistance was related to an overproduction of HGF. These data indicate that MSC source could be of importance while designing immunomodulating cell therapy in transplantation.

PMID: 21700275 [PubMed - indexed for MEDLINE]


Neuronal differentiation of human adipose tissue-derived stem cells for peripheral nerve regeneration in vivo.
Source:  Pub Med
Monday, 29 November 1999 17:00

Neuronal differentiation of human adipose tissue-derived stem cells for peripheral nerve regeneration in vivo.

Arch Surg. 2011 Jun;146(6):666-74

Authors: Scholz T, Sumarto A, Krichevsky A, Evans GR

Abstract
OBJECTIVE: To evaluate the ability of a tissue-engineered nerve construct composed of a nerve guidance channel and neurally differentiated human adipose tissue-derived stem cells (hASCs) to enhance peripheral nerve regeneration in a rat sciatic nerve model.
DESIGN: A 13-mm sciatic nerve gap was bridged with silastic conduits in 64 athymic nude rats, and differentiated hASCs were implanted into the nerve gap. The effect of repetitive renewal of differentiation medium on days 14 and 28 was further tested. Adequate negative controls and isograft controls were used.
SETTING: Academic research.
PATIENTS: The hASCs were isolated from human adipose tissue of patients undergoing liposuction procedures.
MAIN OUTCOME MEASURES: Direct measurements of nerve function included sciatic functional index score, extensor postural thrust, and sensory evaluation. Indirect measurements included gastrocnemius and soleus muscle atrophy. Histomorphometric evaluation included the number and diameter of axons and fibers, nerve fiber density, myelin thickness, g-ratio (axon diameter-total fiber diameter ratio), and myelin thickness-axon diameter ratio.
RESULTS: The use of hASCs demonstrated significantly improved functional recovery as measured by the sciatic functional index, extensor postural thrust, sensory evaluation, and gastrocnemius and soleus muscle weight after 14 days and 1, 2, 3, and 4 months. Groups with their medium renewed also demonstrated further enhanced functional recovery compared with their counterparts that did not have their medium renewed.
CONCLUSION: This tissue-engineered nerve construct using hASCs was able to improve functional recovery during the first 4 months, comparable with nerve isografts.

PMID: 21690442 [PubMed - indexed for MEDLINE]


Adipose-derived stromal cell transplantation for treatment of stress urinary incontinence.
Source:  Pub Med
Monday, 29 November 1999 17:00

Adipose-derived stromal cell transplantation for treatment of stress urinary incontinence.

Tissue Cell. 2011 Aug;43(4):246-53

Authors: Wu G, Song Y, Zheng X, Jiang Z

Abstract
We aimed to investigate the application of adipose-derived stromal cells in the treatment of stress urinary incontinence (SUI). Animal models of stress urinary incontinence were established with Sprague-Dawley female rats by complete cutting of the pudendal nerve. Rat adipose-derived stromal cells were isolated, cultured and successfully transplanted into animal models. Effects of stem cell transplantation were evaluated through urodynamic testing and morphologic changes of the urethra and surrounding tissues before and after transplantation. Main urodynamic outcome measures were measured. Intra-bladder pressure and leak point pressure were measured during filling phase. Morphologic examinations were performed. Transplantation of adipose-derived stem cells significantly strengthened local urethral muscle layers and significantly improved the morphology and function of sphincters. Urodynamic testing showed significant improvements in maximum bladder capacity, abdominal leak point pressure, maximum urethral closure pressure, and functional urethral length. Morphologic changes and significant improvement in urination control were consistent over time. It was concluded that periurethral injection of adipose-derived stromal cells improves function of the striated urethral sphincter, resulting in therapeutic effects on SUI. Reconstruction of the pelvic floor through transplantation of adipose-derived cells is a minimally invasive and effective treatment for SUI.

PMID: 21704350 [PubMed - indexed for MEDLINE]


Harvey Cushing's repair of a dural defect after a traumatic brain injury: novel use of a fat graft.
Source:  Pub Med
Monday, 29 November 1999 17:00

Harvey Cushing's repair of a dural defect after a traumatic brain injury: novel use of a fat graft.

World Neurosurg. 2011 May-Jun;75(5-6):696-9

Authors: Zaidi HA, Pendleton C, Cohen-Gadol AA, Quinones-Hinojosa A

Abstract
OBJECTIVE: A review of Harvey Cushing's surgical cases at Johns Hopkins Hospital revealed new information about his early work with the use of fat grafts to close dural defects.
METHODS: The Johns Hopkins Hospital surgical records from 1896 to 1912 were reviewed. A single case in which Cushing used an autologous fat graft to repair a dural defect in 1912 after a traumatic brain injury was selected for further study.
RESULTS: An 18-year-old white female patient presented with recurring seizures in her sleep approximately 12 years after a traumatic brain injury. A depressed skull fracture as a result of this injury was explored by Cushing, and a dural defect was found. Fat was harvested from the thigh of the patient and was sutured in layers to cover the defect. Cushing noted that this was the first of its kind of operation and "probably of very little use." The patient was discharged within 2 weeks after the operation despite recurrence of her seizure episodes.
CONCLUSIONS: Despite its questionable functional success, we report herein a previously unpublished operative case by Harvey Cushing's in which an autologous fat graft was used to close a traumatic wound resulting in a dural defect. This report predates currently known published reports of the use of fat to seal dural defects. Cushing recognized that a factor found in fat tissue may aid in wound healing. Contemporary studies indicate the presence of mesenchymal stem cells in fat tissue may be responsible for the accelerated healing and reduced incidence of cerebral spinal fluid leaks after cranial surgery.

PMID: 21704938 [PubMed - indexed for MEDLINE]


Once fat was fat and that was that: our changing perspectives on adipose tissue.
Source:  Pub Med
Monday, 29 November 1999 17:00

Once fat was fat and that was that: our changing perspectives on adipose tissue.

Cardiovasc J Afr. 2011 May-Jun;22(3):147-54

Authors: Ferris WF, Crowther NJ

Abstract
Past civilisations saw excess body fat as a symbol of wealth and prosperity as the general population struggled with food shortages and famine. Nowadays it is recognised that obesity is associated with co-morbidities such as cardiovascular disease and diabetes. Our views on the roll of adipose tissue have also changed, from being solely a passive energy store, to an important endocrine organ that modulates metabolism, immunity and satiety. The relationship between increased visceral adiposity and obesity-related co-morbidities has lead to the recognition that variation in fat distribution contributes to ethnic differences in the prevalence of obesity-related diseases. Our current negative view of adipose tissue may change with the use of pluripotent adipose-derived stromal cells, which may lead to future autologous stem cell therapies for bone, muscle, cardiac and cartilage disorders. Here, we briefly review the concepts that adipose tissue is an endocrine organ, that differences in body fat distribution underline the aetiology of obesity-related co-morbidities, and the use of adipose-derived stem cells for future therapies.

PMID: 21713306 [PubMed - indexed for MEDLINE]


Favorable Response of Chronic Refractory Immune Thrombocytopenic Purpura to Mesenchymal Stem Cells.
Source:  Pub Med
Monday, 29 November 1999 17:00

Favorable Response of Chronic Refractory Immune Thrombocytopenic Purpura to Mesenchymal Stem Cells.

Stem Cells Dev. 2011 Aug 12;

Authors: Fang B, Mai L, Li N, Song Y

Abstract
Seven patients with chronic refractory immune thrombocytopenic purpura (ITP) received adipose tissue-derived mesenchymal stem cells (AMSC) from haplo-identical family donors. The AMSC dose was 2.0×10(6)/kg. No side effects were noted after the AMSC infusions. Overall responses were reached in all patients and sustained response rate was 57.1% (4/7). The serum levels of transforming growth factor β1 (TGF-β1), interleukin (IL)-4, and IL-10 were significantly elevated, whereas those of interferon-γ (IFN-γ) and IL-2 were significantly decreased after AMSC administration, compared with those in the patients with active ITP. During follow-up, the cytokine profiles in patients maintaining sustained response remained stable compared with the post-treatment level, but IFN-γ and IL-2 levels were significantly increased, and those of TGF-β1, IL-4, and IL-10 were significantly reduced again in relapsed patients. AMSC therapy seems to represent reasonable salvage treatment in severe, chronic refractory ITP by causing a shift in the Th1/Th2 cytokine balance to the same levels as normal controls.

PMID: 21711157 [PubMed - as supplied by publisher]


Telomere length, telomerase activity and osteogenic differentiation are maintained in adipose-derived stromal cells from senile osteoporotic SAMP6 mice.
Source:  Pub Med
Monday, 29 November 1999 17:00

Telomere length, telomerase activity and osteogenic differentiation are maintained in adipose-derived stromal cells from senile osteoporotic SAMP6 mice.

J Tissue Eng Regen Med. 2011 Jun 28;

Authors: Mirsaidi A, Kleinhans KN, Rimann M, Tiaden AN, Stauber M, Rudolph KL, Richards PJ

Abstract
Adipose tissue provides for a rich and easily accessible source of multipotent stromal cells and thus offers the potential for autologous cell-based therapy for a number of degenerative diseases. Senile osteoporosis is characterized by a reduction in bone quality, which is associated with inadequacies in bone marrow stromal cell (BMSC) differentiation. In the present study, we have characterized adipose-derived stromal cells (ASCs) isolated from aged osteoporotic mice and evaluated their suitability as a source of osteogenic precursor cells. Significant reductions in both tibia bone quality and telomere length in liver tissue were observed in the senescence-accelerated mouse prone 6 strain (SAMP6), as compared to the control age-matched senescence-accelerated mouse resistant 1 strain (SAMR1), thus confirming osteoporosis and accelerated ageing traits in this model. ASCs isolated from inguinal fat expressed mesenchymal surface markers and were capable of differentiating along the osteoblast, adipocyte and chondrocyte lineages. Telomere length was not compromised in ASCs from SAMP6 mice but was actually found to be significantly increased as compared to control SAMR1 mice. Furthermore, ASCs from both strains were comparable in terms of telomerase activity, p21 mRNA expression, SA-β-gal activity and proliferative capacity. The overall osteogenic and adipogenic potential of ASCs was comparable between SAMP6 and SAMR1 strains, as determined by quantitative molecular, biochemical and histological analyses. In conclusion, adipose tissue may represent a promising autologous cell source for the development of novel bone regenerative therapeutic strategies in the treatment of age-related osteoporosis. Copyright © 2011 John Wiley & Sons, Ltd.

PMID: 21710574 [PubMed - as supplied by publisher]


Regeneration of human bones in hip osteonecrosis and human cartilage in knee osteoarthritis with autologous adipose-tissue-derived stem cells: a case series.
Source:  Pub Med
Monday, 29 November 1999 17:00

Regeneration of human bones in hip osteonecrosis and human cartilage in knee osteoarthritis with autologous adipose-tissue-derived stem cells: a case series.

J Med Case Reports. 2011;5(1):296

Authors: Pak J

Abstract
UNLABELLED: ABSTRACT:
INTRODUCTION: This is a series of clinical case reports demonstrating that a combination of percutaneously injected autologous adipose-tissue-derived stem cells, hyaluronic acid, platelet rich plasma and calcium chloride may be able to regenerate bones in human osteonecrosis, and with addition of a very low dose of dexamethasone, cartilage in human knee osteoarthritis.
CASE REPORTS: Stem cells were obtained from adipose tissue of abdominal origin by digesting lipoaspirate tissue with collagenase. These stem cells, along with hyaluronic acid, platelet rich plasma and calcium chloride, were injected into the right hip of a 29-year-old Korean woman and a 47-year-old Korean man. They both had a history of right hip osteonecrosis of the femoral head. For cartilage regeneration, a 70-year-old Korean woman and a 79-year-old Korean woman, both with a long history of knee pain due to osteoarthritis, were injected with stem cells along with hyaluronic acid, platelet rich plasma, calcium chloride and a nanogram dose of dexamethasone. Pre-treatment and post-treatment MRI scans, physical therapy, and pain score data were then analyzed.
CONCLUSIONS: The MRI data for all the patients in this series showed significant positive changes. Probable bone formation was clear in the patients with osteonecrosis, and cartilage regeneration in the patients with osteoarthritis. Along with MRI evidence, the measured physical therapy outcomes, subjective pain, and functional status all improved. Autologous mesenchymal stem cell injection, in conjunction with hyaluronic acid, platelet rich plasma and calcium chloride, is a promising minimally invasive therapy for osteonecrosis of femoral head and, with low-dose dexamethasone, for osteoarthritis of human knees.

PMID: 21736710 [PubMed - in process]


Immunological Aspects of Allogeneic and Autologous Mesenchymal Stem Cell Therapies.
Source:  Pub Med
Monday, 29 November 1999 17:00

Immunological Aspects of Allogeneic and Autologous Mesenchymal Stem Cell Therapies.

Hum Gene Ther. 2011 Aug 10;

Authors: Hoogduijn MJ, Roemeling-van Rhijn M, Korevaar SS, Engela AU, Weimar W, Baan CC

Abstract
Abstract Mesenchymal stem cells (MSCs) have potential for therapeutic application as an immunomodulatory and regenerative agent. The immunogenicity and survival of MSCs after infusion are, however, not clear and evidence suggests that allogeneic but also autologous MSCs disappear rapidly after infusion. This may be associated with the susceptibility of MSCs to lysis by natural killer (NK) cells, possibly a result of culture-induced stress. In the present study we examined whether NK cell-mediated lysis of MSCs could be inhibited by immunosuppressive drugs. Human MSCs were isolated from adipose tissue and expanded in culture. Peripheral blood mononuclear cells were activated with interleukin (IL)-2 (200 U/ml) and IL-15 (10 ng/ml) for 7 days. CD3(-)CD16(+)CD56(+) NK cells were then isolated by fluorescence-activated cell sorting and added to europium-labeled MSCs for 4 hr in the presence or absence of immunosuppressive drugs. Lysis of MSCs was determined by spectrophotometric measurement of europium release. Nonactivated NK cells were not capable of lysing MSCs. Cytokine-activated NK cells showed upregulated levels of granzyme B and perforin and efficiently lysed allogeneic and autologous MSCs. Addition of tacrolimus, rapamycin or sotrastaurin to the lysis assay did not inhibit MSC killing. Furthermore, preincubation of activated NK cells with the immunosuppressive drugs for 24 hr before exposure to MSCs had no effect on MSC lysis. Last, addition of the immunosuppressants before and during the activation of NK cells, reduced NK cell numbers but did not affect their capacity to lyse MSCs. We conclude that the immunosuppressive drugs tacrolimus, rapamycin, and sotrastaurin are not capable of inhibiting the lysis of allogeneic and autologous MSCs by activated NK cells. Other approaches to controlling lysis of MSCs should be investigated, as controlling lysis may determine the efficacy of MSC therapy.

PMID: 21732766 [PubMed - as supplied by publisher]


Human adipose tissue-derived mesenchymal stem cells expressing yeast cytosinedeaminase::uracil phosphoribosyltransferase inhibit intracerebral rat glioblastoma.
Source:  Pub Med
Monday, 29 November 1999 17:00

Human adipose tissue-derived mesenchymal stem cells expressing yeast cytosinedeaminase::uracil phosphoribosyltransferase inhibit intracerebral rat glioblastoma.

Int J Cancer. 2011 Jul 5;

Authors: Altanerova V, Cihova M, Babic M, Rychly B, Ondicova K, Mravec B, Altaner C

Abstract
Prodrug cancer gene therapy by mesenchymal stem cells targeted to tumors represents an attractive tool to activate prodrugs directly within the tumor mass, thus avoiding systemic toxicity. In this study, we tested the feasibility and efficacy of human adipose tissue-derived mesenchymal stem cells, engineered to express the suicide gene cytosine deaminase::uracil phosphoribosyltransferase to treat intracranial rat C6 glioblastoma. Experiments were designed to simulate conditions of future clinical application for high-grade glioblastoma therapy by direct injections of therapeutic stem cells into tumor. We demonstrated that genetically modified therapeutic stem cells still have the tumor tropism when injected to a distant intracranial site and effectively inhibited glioblastoma growth after 5-FC therapy. Co-administration of C6 cells and therapeutic stem cells with delayed 5-FC therapy improved the survival in a therapeutic stem cell dose-dependent manner and induced complete tumor regression in a significant number of animals. Continuous intracerebro-ventricular delivery of 5-FC using osmotic pump reduced the dose of prodrug required for the same therapeutic effect, and along with repeated administration of therapeutic stem cells increased the survival time. Intracerebral injection of therapeutic stem cells and treatment with 5-FC did not show any detectable adverse effects. Results support the arguments to begin clinical studies for treatment of high-grade brain tumors.

PMID: 21732344 [PubMed - as supplied by publisher]


Basic science review on adipose tissue for clinicians.
Source:  Pub Med
Monday, 29 November 1999 17:00

Basic science review on adipose tissue for clinicians.

Plast Reconstr Surg. 2011 Sep;128(3):829-30

Authors: Klinger FM, Vinci V, Forcellini D, Caviggioli F

PMID: 21866031 [PubMed - indexed for MEDLINE]


Harnessing the therapeutic potential of mesenchymal stem cells in multiple sclerosis.
Source:  Pub Med
Monday, 29 November 1999 17:00

Harnessing the therapeutic potential of mesenchymal stem cells in multiple sclerosis.

Expert Rev Neurother. 2011 Sep;11(9):1295-303

Authors: Darlington PJ, Boivin MN, Bar-Or A

Abstract
Phase I clinical trials exploring the use of autologous mesenchymal stem cell (MSC) therapy for the treatment of multiple sclerosis (MS) have begun in a number of centers across the world. MS is a complex and chronic immune-mediated and neurodegenerative disease influenced by genetic susceptibility and environmental risk factors. The ideal treatment for MS would involve both attenuation of detrimental inflammatory responses, and induction of a degree of tissue protection/regeneration within the CNS. Preclinical studies have demonstrated that both human-derived and murine-derived MSCs are able to improve outcomes in the animal model of MS, experimental autoimmune encephalomyelitis. How MSCs ameliorate experimental autoimmune encephalomyelitis is being intensely investigated. One of the major mechanisms of action of MSC therapy is to inhibit various components of the immune system that contribute to tissue destruction. Emerging evidence now supports the idea that MSCs can access the CNS where they can provide protection against tissue damage, and may facilitate tissue regeneration through the production of growth factors. The prospect of cell-based therapy using MSCs has several advantages, including the relative ease with which they can be extracted from autologous bone marrow or adipose tissue and expanded in vitro to reach the purity and numbers required for transplantation, and the fact that MSC therapy has already been used in other human disease settings, such as graft-versus-host and cardiac disease, with initial reports indicating a good safety profile. This article will focus on the theoretical and practical issues relevant to considerations of MSC therapy in the context of MS.

PMID: 21864075 [PubMed - in process]


Human Adipose-Derived Stem Cells Impair Natural Killer Cell Function and Exhibit Low Susceptibility to Natural Killer-Mediated Lysis.
Source:  Pub Med
Monday, 29 November 1999 17:00

Human Adipose-Derived Stem Cells Impair Natural Killer Cell Function and Exhibit Low Susceptibility to Natural Killer-Mediated Lysis.

Stem Cells Dev. 2011 Oct 18;

Authors: Delarosa O, Sánchez-Correa B, Morgado S, Ramírez C, Del Río B, Menta R, Lombardo E, Tarazona R, Casado JG

Abstract
Human adipose-derived stem cells (hASCs) have been successfully used in treating numerous diseases. However, several aspects need to be considered, particularly in the context of allogeneic cell therapy. To better understand hASCs-host interactions, we studied the phenotype of hASCs and their modulatory effect on natural killer (NK) cells by using bone marrow-mesenchymal stem cells (hBM-MSCs) as a reference. The hASCs displayed a lower susceptibility to NK cell-mediated lysis and a lower expression of ligands for DNAM-1 when compared with hBM-MSCs. Moreover, here we demonstrated that hASCs and hBM-MSCs can modulate NK cells through the action of soluble factors such as indoleamine 2,3-dioxygenase. Altogether, these results suggest that for an adoptive cell therapy based on the transfer of allogeneic hASCs, the NK-hASCs crosstalk will not result in an immediate recognition of the transferred cells. Thus, hASCs may remain in the tissue long enough to balance the immune response before being cleared.

PMID: 21867426 [PubMed - as supplied by publisher]


In vitro expansion of human adipose-derived stem cells in a spinner culture system using human extracellular matrix powders.
Source:  Pub Med
Monday, 29 November 1999 17:00

In vitro expansion of human adipose-derived stem cells in a spinner culture system using human extracellular matrix powders.

Cell Tissue Res. 2011 Sep;345(3):415-23

Authors: Choi JS, Kim BS, Kim JD, Choi YC, Lee EK, Park K, Lee HY, Cho YW

Abstract
Stem cell therapy requires large numbers of stem cells to replace damaged tissues, but only limited numbers of stem cells can be harvested from a single patient. To obtain large quantities of stem cells with differentiation potential, we explored a spinner culture system using human extracellular matrix (hECM) powders. The hECM was extracted from adipose tissue and fabricated into powders. Human adipose-derived stem cells (hASCs) were isolated, seeded on hECM powders, and cultivated in a spinner flask. The 3-D culture system, using hECM powders, was highly effective for promoting cell proliferation. The number of hASCs in the 3-D culture system significantly increased for 10 days, resulting in an approximately 10-fold expansion, whereas a traditional 2-D culture system showed just a 2.8-fold expansion. Surface markers, transcriptional factors, and differentiation potential of hASCs were assayed to identify the characteristics of proliferated cells in 3-D culture system. The hASCs expressed the pluripotency markers, Oct-4 and Sox-2 during 3-D culture and retained their capacity to differentiate into adipogenic, osteogenic, and chondrogenic lineages. These findings demonstrate that the 3-D culture systems using hECM powders provide an efficient in vitro environment for stem cell proliferation, and could act as stem cell delivery carriers for autologous tissue engineering and cell therapy.

PMID: 21866312 [PubMed - in process]


Sources of adult mesenchymal stem cells applicable for musculoskeletal applications - a systematic review of the literature.
Source:  Pub Med
Monday, 29 November 1999 17:00

Sources of adult mesenchymal stem cells applicable for musculoskeletal applications - a systematic review of the literature.

Open Orthop J. 2011;5 Suppl 2:242-8

Authors: Mafi R, Hindocha S, Mafi P, Griffin M, Khan WS

Abstract
Mesenchymal stem cells (MSCs) were first discovered by Friedenstein and his colleagues in 1976 from bone marrow. The unique property of these cells was their potential to develop into fibroblastic colony forming cells. Since Friedenstein's discovery of these cells the interest in adult MSCs has been progressively growing. Nowadays MSCs are defined as undeveloped biological cells capable of proliferation, self renewal and regenerating tissues. All these properties of MSCs have been discovered in the past 35 years. MSCs can play a crucial role in tissue engineering, organogenesis, gene therapy, transplants as well as tissue injuries. These cells were mainly extracted from bone marrow but there have been additional sources for MSCs discovered in the laboratories including: muscle, dermis, trabecular bone, adipose tissue, periosteum, pericyte, blood, synovial membrane and so forth. The discovery of the alternative sources of MSCs helps widen the application of these cells in different areas of medicine. By way of illustration, they can be used in various therapeutic purposes such as tissue regeneration and repair in musculoskeletal diseases including osteonecrosis of femoral head, stimulating growth in children with osteogenesis imperfecta, disc regeneration, osteoarthritis and duchenne muscular dystrophy. In order to fully comprehend the characteristics and potential of MSCs future studies in this field are essential.

PMID: 21886689 [PubMed - in process]


Toward a Clinical-Grade Expansion of Mesenchymal Stem Cells from Human Sources: A Microcarrier-Based Culture System Under Xeno-Free Conditions.
Source:  Pub Med
Monday, 29 November 1999 17:00

Toward a Clinical-Grade Expansion of Mesenchymal Stem Cells from Human Sources: A Microcarrier-Based Culture System Under Xeno-Free Conditions.

Tissue Eng Part C Methods. 2011 Sep 6;

Authors: Santos FD, Andrade PZ, Abecasis MM, Gimble JM, Chase LG, Campbell AM, Boucher S, Vemuri MC, Silva CL, Cabral JM

Abstract
The immunomodulatory properties of mesenchymal stem cells (MSCs) make them attractive therapeutic agents for a wide range of diseases. However, the highly demanding cell doses used in MSC clinical trials (up to millions of cells/kg patient) currently require labor intensive methods and incur high reagent costs. Moreover, the use of xenogenic (xeno) serum-containing media represents a risk of contamination and raises safety concerns. Bioreactor systems in combination with novel xeno-free medium formulations represent a viable alternative to reproducibly achieve a safe and reliable MSC doses relevant for cell therapy. The main goal of the present study was to develop a complete xeno-free microcarrier-based culture system for the efficient expansion of human MSC from two different sources, human bone marrow (BM), and adipose tissue. After 14 days of culture in spinner flasks, BM MSC reached a maximum cell density of (2.0±0.2)×10(5) cells·mL(-1) (18±1-fold increase), whereas adipose tissue-derived stem cells expanded to (1.4±0.5)×10(5) cells·mL(-1) (14±7-fold increase). After the expansion, MSC expressed the characteristic markers CD73, CD90, and CD105, whereas negative for CD80 and human leukocyte antigen (HLA)-DR. Expanded cells maintained the ability to differentiate robustly into osteoblast, adipocyte, and chondroblast lineages upon directed differentiation. These results demonstrated the feasibility of expanding human MSC in a scalable microcarrier-based stirred culture system under xeno-free conditions and represent an important step forward for the implementation of a Good Manufacturing Practices-compliant large-scale production system of MSC for cellular therapy.

PMID: 21895491 [PubMed - as supplied by publisher]


Immune response to human embryonic stem cell-derived cardiac progenitors and adipose-derived stromal cells.
Source:  Pub Med
Monday, 29 November 1999 17:00

Immune response to human embryonic stem cell-derived cardiac progenitors and adipose-derived stromal cells.

J Cell Mol Med. 2011 Sep 5;

Authors: Calderon D, Planat-Benard V, Bellamy V, Vanneaux V, Khun C, Peyrard S, Larghero J, Desnos M, Casteilla L, Pucéat M, Menasché P, Chatenoud L

Abstract
Background. Transplantation of allogeneic human embryonic stem cell-derived cardiac progenitors triggers an immune response. We assessed whether this response could be modulated by the concomitant use of adipose-derived stromal cells (ADSC). Methods. Peripheral blood mononuclear cells were collected from 40 patients with coronary artery disease (CAD) and 9 healthy controls. Cardiac progenitors (CD15(+) Mesp1(+) ) were generated as already reported from the I6 cell line treated with bone morphogenetic protein (BMP)-2. ADSC were obtained from abdominal dermolipectomies. We assessed the proliferative response of peripheral lymphocytes from patients and controls to cardiac progenitors cultured on a monolayer of ADSC, to allogeneic lymphocytes in mixed lymphocyte culture and to the T cell mitogen phytohemaglutin A (PHA) in presence or absence of ADSC. Results. Cardiac progenitors cultured on a monolayer of ADSC triggered a proliferation of lymphocytes from both patients and controls albeit lower than that induced by allogeneic lymphocytes. When cultured alone, ADSC did not induce any proliferation of allogeneic lymphocytes. When added to cultures of lymphocytes, ADSC significantly inhibited the alloantigen or mitogen-induced proliferative response. Compared to healthy controls, lymphocytes from patients presenting CAD expressed a decreased proliferative capacity, in particular to mitogen-induced stimulation. Conclusion. ADSC express an immunomodulatory effect that limits both alloantigen and mitogen-induced lymphocyte responses. Furthermore, lymphocytes from patients with CAD are low responders to conventional stimuli, possibly because of their age and disease-associated treatment regimens. We propose that, in combination, these factors may limit the in vivo immunogenicity of cardiac progenitors co-implanted with ADSC in patients with CAD.

PMID: 21895965 [PubMed - as supplied by publisher]


[Correlation between hypertension and erectile dysfunction].
Source:  Pub Med
Monday, 29 November 1999 17:00

[Correlation between hypertension and erectile dysfunction].

Zhonghua Nan Ke Xue. 2011 Aug;17(8):675-81

Authors: Liu JH, Ling Q

Abstract
The relationship between erectile dysfunction (ED) and hypertension is a focus in andrological research. ED and hypertension share some pathophysiologic pathways, such as oxidative stress-induced endothelial dysfunction and up-regulated RhoA/Rho kinase activity, and both are the diseases at different stages of the pathological process of vascular dysfunction. Thus, it is particularly important to conduct regular and meticulous evaluation of such patients, so as to give rational individualized medication. Phosphodiesterase-5 inhibitors have an excellent efficacy and safety profile in the management of hypertension, either used alone or with antihypertensive medication. At present, gene therapy and adipose-derived stem cell therapy have displayed favorable prospects in the management of ED and hypertension, and translational medicine may help bring more clinical benefits.

PMID: 21898987 [PubMed - in process]


Production of canine mesenchymal stem cells from adipose tissue and application in dogs with chronic osteoarthritis of the humeroredal joints.
Source:  Pub Med
Monday, 29 November 1999 17:00

Production of canine mesenchymal stem cells from adipose tissue and application in dogs with chronic osteoarthritis of the humeroredal joints.

Cell Biol Int. 2011 Sep 22;

Authors: Guercio A, Di Marco P, Casella S, Cannella V, Russotto L, Purpari G, Di Bella S, Piccione G

Abstract
Autologous adipose-derived mesenchymal stem cell (AD-MSC) therapy involves harvesting fat from the patient isolating the stem and regenerative cells and administering the cells back to the patient. The aim of this study was to evaluate the production of canine AD-MSCs and the possible application of cellular therapy in dogs. In order to assess whether cellular therapy can replace the drug therapy, the clinical effect of a single intraarticular injection of AD-MSCs was evaluated on four dogs with lameness associated with osteoarthritis (OA) of humeroradial joints. The study demonstrated that the mesenchymal stem cells (MSCs) can be readily isolated in adult dog from adipose tissue confirming their ability to form colony and to differentiate into a variety of cell phenotypes. AD-MSCs expressed OCT4, NANOG and SOX2 at mRNA level, pluripotency markers usually ascribed to embryonic stem cells. These results suggested the stemness of the cells isolated from canine fat and good results of quality controls ensured the availability of AD-MSCs for clinical and experimental use. Follow-up studies, designed to evaluate the effects of AD-MSC therapy, showed that treated dogs' osteoarthritis of the elbow joints improved over time. In conclusion, the cumulative data indicated that this cellular therapy shows a significant potential for clinical use in the treatment of lameness associated with osteoarthritis. Considering that cellular therapy showed substantial promise in the treatment of osteoarthritis it should be applied before the injury becomes severe.

PMID: 21936851 [PubMed - as supplied by publisher]


Detection of embryonic stem cell markers in adult human adipose tissue-derived stem cells.
Source:  Pub Med
Monday, 29 November 1999 17:00

Detection of embryonic stem cell markers in adult human adipose tissue-derived stem cells.

Indian J Pathol Microbiol. 2011 Jul-Sep;54(3):501-8

Authors: Arumugam SB, Trentz OA, Arikketh D, Senthinathan V, Rosario B, Mohandas PV

Abstract
BACKGROUND: Bone marrow transplantation is already an established therapy, which is now widely used in medicine to treat leukemia, lymphoma, and several inherited blood disorders. The culture of multilineage cells from easily available adipose tissue is another source of multipotent mesenchymal stem cells, and is referred to as adipose tissue-derived stem cells (ADSCs). While ADSCs are being used to treat various conditions, some lacuna exists regarding the specific proteins in these. It was therefore decided to analyze the specific proteins of embryonic cells in ADSCs.
AIMS: To analyze the specific protein of embryonic stem cells (ESCs) in ADSCs.
MATERIALS AND METHODS: Adult human adipose tissue-derived stem cells (ADSCs) were harvested from 13 patients after obtaining patients' consent. The specific markers of ESCs included surface proteins CD10, CD13, CD44, CD59, CD105, and CD166, and further nucleostemin, (NS) NANOG, peroxisome proliferator-activated receptor-gγ, collagen type 1 (Coll1), alkaline phosphate, (ALP) osteocalcin (OC), and core binding factor 1 (Cbfa1) were analyzed using by reverse transcription-polymerase chain reaction, (RT-PCR) immunofluorescence (IF), and western blot.
RESULTS: All the proteins were expressed distinctly, except CD13 and OC. CD13 was found individually with different expressions, and OC expression was discernable.
CONCLUSIONS: Although the ESC with its proven self-renewal capacity and pluripotency seems appropriate for clinical use, the recent work on ADSCs suggests that these adult stem cells would be a valuable source for future biotechnology, especially since there is a relative ease of procurement.

PMID: 21934210 [PubMed - in process]


Progress in stem cell biology in regenerative medicine for liver disease.
Source:  Pub Med
Monday, 29 November 1999 17:00

Progress in stem cell biology in regenerative medicine for liver disease.

Hepatol Res. 2011 Sep 22;

Authors: Shiota G, Yasui T

Abstract
Regenerative medicine using stem cells has attracted much attention, since stem cells are responsible for highly proliferative activity and multipotential ability of differentiation. Induced pluripotent stem cells and embryonic stem cells or the adult stem cells such as bone marrow-derived stem cells and adipose tissue-derived stem cells have been expected as a cell source of regenerative medicine. Since differentiating methods of human stem cells into the defined lineage of cells remains to be developed, we focus on the differentiating strategies of pluripotent stem cells and mesenchymal stem cells into liver lineage, especially on cytokine function and gene expression during hepatic differentiation. The survey of previously published papers discloses that the protocols that mimic the liver developmental process seem to be effective in obtaining functional hepatocytes. However, in order to develop hepatic regenerative medicine that is useful in a clinical setting, more effective and potent strategies that obtain mature hepatocytes are required.

PMID: 21951276 [PubMed - as supplied by publisher]


Estimation of the Distribution of Intravenously Injected Adipose Tissue-Derived Stem Cells Labeled with Quantum Dots in Mice Organs through the Determination of their Metallic Components by ICPMS.
Source:  Pub Med
Monday, 29 November 1999 17:00

Estimation of the Distribution of Intravenously Injected Adipose Tissue-Derived Stem Cells Labeled with Quantum Dots in Mice Organs through the Determination of their Metallic Components by ICPMS.

Anal Chem. 2011 Nov 1;83(21):8252-8

Authors: Takasaki Y, Watanabe M, Yukawa H, Sabarudin A, Inagaki K, Kaji N, Okamoto Y, Tokeshi M, Miyamoto Y, Noguchi H, Umemura T, Hayashi S, Baba Y, Haraguchi H

Abstract
Adipose tissue-derived stem cells (ASCs) have shown promise in cell therapy because of their ability to self-renew damaged or diseased organs and easy harvest. To ensure the distribution and quantification of the ASCs injected from tail vein, several whole-body imaging techniques including fluorescence optical imaging with quantum dots (QDs) have been employed, but they may suffer from insufficient sensitivity and accuracy. Here, we report quantitative distribution of ASCs in various organs (heart, lung, liver, spleen, and kidney) of mice, which were intravenously injected with QDs-labeled ASCs (QDs-ASCs), through the detection of QDs-derived metallic components by inductively coupled plasma mass spectrometry (ICPMS). For accurate and precise determination, each organ was harvested and completely digested with a mixture of HNO(3) and H(2)O(2) in a microwave oven prior to ICPMS measurement, which was equipped with a microflow injection system and a laboratory-made capillary-attached micronebulizer. After optimization, 16 elements including major components (Cd, Se, and Te) of QDs and essential elements (Na, K, Mg, Ca, P, S, Mn, Fe, Co, Cu, Zn, Se, Sr, and Mo) were successfully determined in the organs. As compared to untreated mice, QDs-ASCs-treated mice showed significantly higher levels of Cd and Te in all organs, and as expected, the molar ratio of Cd to Te in each organ was in good agreement with the molar composition ratio in the QDs. This result indicates that the increment of Cd (or Te) can be used as a tracer for calculating the distribution of ASCs in mice organs. As a result of the calculation, 36.8%, 19.1%, 0.59%, 0.49%, and 0.25% of the total ASCs injected were estimated to be distributed in the liver, lung, heart, spleen, and kidney, respectively.

PMID: 21958307 [PubMed - in process]


[Stem cell properties of therapeutic potential].
Source:  Pub Med
Monday, 29 November 1999 17:00

[Stem cell properties of therapeutic potential].

Korean J Gastroenterol. 2011 Sep 25;58(3):125-32

Authors: Seo GS

Abstract
Stem cell research is a innovative technology that focuses on using undifferentiated cells able to self-renew through the asymmetrical or symmetrical divisions. Three types of stem cells have been studied in laboratory including embryonic stem cell, adult stem cells and induced pluripotent stem cells. Embryonic stem cells are pluripotent stem cells derived from the inner cell mass and it can give rise to any fetal or adult cell type. Adult stem cells are multipotent, have the ability to differentiate into a limited number of specialized cell types, and have been obtained from the bone marrow, umbilical cord blood, placenta and adipose tissue. Stem cell therapy is the most promising therapy for several degenerative and devastating diseases including digestive tract disease such as liver failure, inflammatory bowel disease, Celiac sprue, and pancreatitis. Further understanding of biological properties of stem cells will lead to safe and successful stem cell therapies. (Korean J Gastroenterol 2011;58: 125-132).

PMID: 21960099 [PubMed - in process]


[Clinical trials with stem cells in digestive diseases and future perspectives].
Source:  Pub Med
Monday, 29 November 1999 17:00

[Clinical trials with stem cells in digestive diseases and future perspectives].

Korean J Gastroenterol. 2011 Sep 25;58(3):139-43

Authors: Kim TI

Abstract
Many techniques for isolation, expansion and handling of stem cells are being developed rapidly, and preclinical evidence has shown the possibility to use this technology for refractory diseases in the near future. Among refractory digestive diseases, Crohn's disease and liver cirrhosis may be two main diseases where stem cell therapy can be applied for anti-inflammation and regeneration of tissue. Currently, with respect to these two diseases, clinical trials using hematopoietic stem cells and mesenchymal stem cells from bone marrow or adipose tissue have shown some evidence of clinical benefits to immune modulation, suppression of inflammation and regeneration of functional cells. However, for the development of practical stem cell therapy, we need more data on underlying mechanisms, effective subpopulation of stem cells and its sources, and effective parameters for monitoring and estimation. With technical advances, the research on embryonic and induced pluripotent stem cells will also contribute to the new therapeutic strategies for digestive regenerative medicine. In the future, a variety of stem cell therapies may be therapeutic options for refractory digestive diseases, but many technical challenges remain to be solved. (Korean J Gastroenterol 2011;58:139-143).

PMID: 21960101 [PubMed - in process]


Molecular Analysis of the Differentiation Potential of Murine Mesenchymal Stem Cells from Tissues of Endodermal or Mesodermal Origin.
Source:  Pub Med
Monday, 29 November 1999 17:00

Molecular Analysis of the Differentiation Potential of Murine Mesenchymal Stem Cells from Tissues of Endodermal or Mesodermal Origin.

Stem Cells Dev. 2011 Oct 4;

Authors: Viero C, Camassola M, Bellagamba B, Ikuta N, Christoff AP, Meirelles LD, Ayres R, Margis R, Nardi NB

Abstract
Mesenchymal stem cells (MSC) have received great attention due to their remarkable regenerative, angiogenic, anti-apoptotic and immunosuppressive properties. Although conventionally isolated from the bone marrow, they are known to exist in all tissues and organs, raising the question on whether they are identical cell populations or have important differences at the molecular level. To better understand the relationship between MSC resident in different tissues, we analyzed the expression of genes related to pluripotency (SOX2 and OCT-4) and to adipogenic (C/EBP and ADIPOR1), osteogenic (OMD and ALP) and chondrogenic (COL10A1 and TRPV4) differentiation in cultures derived from murine endodermal (lung) and mesodermal (adipose) tissue maintained in different conditions. MSC were isolated from lungs (L-MSC) and inguinal adipose tissue (A-MSC) and cultured in normal conditions, in overconfluence or in inductive medium for osteogenic, adipogenic or chondrogenic differentiation. Cultures were characterized for morphology, immunophenotype and by quantitative real-time RT-PCR for expression of pluripotency genes or markers of differentiation. Bone marrow-derived MSC were also analyzed for comparison of these parameters. L-MSC and A-MSC exhibited the typical morphology, immunophenotype and proliferation and differentiation pattern of mesenchymal stem cells. The analysis of gene expression showed a higher potential of adipose tissue-derived MSC towards the osteogenic pathway and of lung-derived MSC to chondrogenic differentiation, representing an important contribution for the definition of the type of cell to be used in clinical trials of cell therapy and tissue engineering.

PMID: 21970410 [PubMed - as supplied by publisher]


Differentiation potential of human mesenchymal stem cells derived from adipose tissue and bone marrow to sinus node-like cells.
Source:  Pub Med
Monday, 29 November 1999 17:00

Differentiation potential of human mesenchymal stem cells derived from adipose tissue and bone marrow to sinus node-like cells.

Mol Med Report. 2012 Jan;5(1):108-13

Authors: Yang J, Song T, Wu P, Chen Y, Fan X, Chen H, Zhang J, Huang C

Abstract
Adult mesenchymal stem cells (MSCs) hold great promise for the repair of heart defects. Both bone marrow-derived mesenchymal stem cells (BMSCs) and adipose tissue-derived stem cells (ASCs) are multipotent and may be induced by 5-azacytidine to differentiate into cardiomyocytes. However, the differentiation potential of human MSCs into sinus node-like cells has not been studied extensively. The aim of this study was to analyze the differences in proliferation and phenotype of ASCs and BMSCs from the same donors and to evaluate their capacity to differentiate into sinus node-like cells in vitro. Five passaged cells from bone marrow and adipose tissue were treated with 10 µM 5-azacytidine for 48 h and further cultured in complete medium for 4 weeks. A comparative study of cultured ASCs and BMSCs was carried out, and the morphological parameters, proliferative capacity, expression of surface markers and differentiation potential to sinus node-like cells were characterized. No morphologic differences were observed between ASCs and BMSCs. Flow cytometric analysis revealed that ASCs and BMSCs both expressed CD29, CD44, CD90 and CD105 and did not express CD34 and CD14, while CD49d, CD106 and CD34 were differentially expressed. Growth curves and doubling time determined with the Cell Counting Kit-8 (CCK-8) demonstrated that ASCs had a stronger proliferative ability than BMSCs. Histological immuofluorescence staining suggested that ASCs and BMSCs were capable of differentiating into sinus node-like cells and that the positive expression ratios of cTNI were higher in ASCs compared to BMSCs at 4 weeks. Expression of the HCN2 and HCN4 genes was detected by reverse transcriptase polymerase chain reaction, and the results revealed that the expression of the HCN genes appeared earlier in ASC-derived sinus node-like cells. ASCs expressed HCN2 and HCN4 shortly after induction with 5-azacytidine for 2 weeks, although BMSCs expressed these genes after 4 weeks. The expression levels of HCN2 and HCN4 mRNA in ASC-derived cells were higher compared to those of BMSCs at 4 weeks. In conclusion, ASCs may be a better candidate as a novel source of cell therapy in sinus bradycardia disorders than BMSCs.

PMID: 21971826 [PubMed - in process]


Stem cells: novel players in the treatment of erectile dysfunction.
Source:  Pub Med
Monday, 29 November 1999 17:00

Stem cells: novel players in the treatment of erectile dysfunction.

Asian J Androl. 2011 Oct 17;

Authors: Zhang H, Albersen M, Jin X, Lin G

Abstract
Stem cells are defined by their capacity for both self-renewal and directed differentiation; thus, they represent great promise for regenerative medicine. Historically, stem cells have been categorized as either embryonic stem cells (ESCs) or adult stem cells (ASCs). It was previously believed that only ESCs hold the ability to differentiate into any cell type, whereas ASCs have the capacity to give rise only to cells of a given germ layer. More recently, however, numerous studies demonstrated the ability of ASCs to differentiate into cell types beyond their tissue origin. The aim of this review was to summarize contemporary evidence regarding stem cell availability, differentiation, and more specifically, the potential of these cells in the diagnosis and treatment of erectile dysfunction (ED) in both animal models and human research. We performed a search on PubMed for articles related to definition, localisation and circulation of stem cells as well as the application of stem cells in both diagnosis and treatment of ED. Strong evidence supports the concept that stem cell therapy is potentially the next therapeutic approach for ED. To date, a large spectrum of stem cells, including bone marrow mesenchymal stem cells, adipose tissue-derived stem cells and muscle-derived stem cells, have been investigated for neural, vascular, endothelial or smooth muscle regeneration in animal models for ED. In addition, several subtypes of ASCs are localized in the penis, and circulating endogenous stem cells can be employed to predict the outcome of ED and ED-related cardiovascular diseases.Asian Journal of Andrology advance online publication, 17 October 2011; doi:10.1038/aja.2011.79.

PMID: 22002437 [PubMed - as supplied by publisher]


Improving viability of stem cells during syringe needle flow through the design of hydrogel cell carriers.
Source:  Pub Med
Monday, 29 November 1999 17:00

Improving viability of stem cells during syringe needle flow through the design of hydrogel cell carriers.

Tissue Eng Part A. 2011 Oct 19;

Authors: Aguado B, Mulyasasmita W, Su J, Lampe KJ, Heilshorn S

Abstract
Cell transplantation is a promising therapy for a myriad of debilitating diseases; however, current delivery protocols using direct injection result in poor cell viability. We demonstrate that during the actual cell injection process, mechanical membrane disruption results in significant acute loss of viability at clinically relevant injection rates. As a strategy to protect cells from these damaging forces, we hypothesize that cell encapsulation within hydrogels of specific mechanical properties will significantly improve viability. We use a controlled in vitro model of cell injection to demonstrate success of this acute protection strategy for a wide range of cell types including human umbilical vein endothelial cells (HUVEC), human adipose stem cells (hASC), rat mesenchymal stem cells (rMSC), and mouse neural progenitor cells (mNPC). Specifically, alginate hydrogels with plateau storage moduli (G') ranging from 0.33 to 58.1 Pa were studied. A compliant crosslinked alginate hydrogel (G'=29.6 Pa) yielded the highest HUVEC viability, 88.9±5.0%, while Newtonian solutions (i.e., buffer only) resulted in 58.7±8.1% viability. Either increasing or decreasing the hydrogel storage modulus reduced this protective effect. Furthermore, cells within non-crosslinked alginate solutions had viabilities lower than media alone, demonstrating that the protective effects are specifically a result of mechanical gelation and not the biochemistry of alginate. Experimental and theoretical data suggest that extensional flow at the entrance of the syringe needle is the main cause of acute cell death. These results provide mechanistic insight into the role of mechanical forces during cell delivery and support the use of protective hydrogels in future clinical stem cell injection studies.

PMID: 22011213 [PubMed - as supplied by publisher]


Current Clinical Therapies for Cartilage Repair, their Limitation and the Role of Stem Cells.
Source:  Pub Med
Monday, 29 November 1999 17:00

Current Clinical Therapies for Cartilage Repair, their Limitation and the Role of Stem Cells.

Curr Stem Cell Res Ther. 2011 Oct 17;

Authors: Dhinsa BS, Adesida AB

Abstract
The management of osteochondral defects of articular cartilage, whether from trauma or degenerative disease, continues to be a significant challenge for Orthopaedic surgeons. Current treatment options such as abrasion arthroplasty procedures, osteochondral transplantation and autologous chondrocyte implantation fail to produce repair tissue exhibiting the same mechanical and functional properties of native articular cartilage. This results in repair issue that inevitably fails as it is unable to deal with the mechanical demands of articular cartilage, and does not prevent further degeneration of the native cartilage. Mesenchymal stem cells have been proposed as a potential source of cells for cell-based cartilage repair due to their ability to self-renew and undergo multi-lineage differentiation. This proposed procedure has the advantage of not requiring harvesting of cells from the joint surface, and its associated donor site morbidity, as well as having multiple possible adult donor tissues such as bone marrow, adipose tissue and synovium. Mesenchymal stem cells have multi-lineage potential, but can be stimulated to undergo chondrogenesis in the appropriate culture medium. As the majority of work with mesenchymal stem cell-derived articular cartilage repair has been carried out in vitro and in animal studies, more work still has to be done before this technique can be used for clinical purposes. This includes realizing the ideal method of harvesting mesenchymal stem cells, the culture medium to stimulate proliferation and differentiation, appropriate choice of scaffold incorporating growth factors directly or with gene therapy and integration of repair tissue with native tissue.

PMID: 22023635 [PubMed - as supplied by publisher]


Precipitation of nanohydroxyapatite on PLLA/PBLG/Collagen nanofibrous structures for the differentiation of adipose derived stem cells to osteogenic lineage.
Source:  Pub Med
Monday, 29 November 1999 17:00

Precipitation of nanohydroxyapatite on PLLA/PBLG/Collagen nanofibrous structures for the differentiation of adipose derived stem cells to osteogenic lineage.

Biomaterials. 2011 Oct 31;

Authors: Ravichandran R, Venugopal JR, Sundarrajan S, Mukherjee S, Ramakrishna S

Abstract
Tissue engineering and nanotechnology have enabled engineering of nanostructured materials to meet the current challenges in bone treatment owing to rising occurrence of bone diseases, accidental damages and defects. Poly(l-lactic acid)/Poly-benzyl-l-glutamate/Collagen (PLLA/PBLG/Col) scaffolds were fabricated by electrospinning and nanohydroxyapatite (n-HA) was deposited by calcium-phosphate dipping method for bone tissue engineering (BTE). The abundance and accessibility of adipose derived stem cells (ADSC) may prove to be novel cell therapeutics for bone repair and regeneration. ADSCs were cultured on these scaffolds and were induced to undergo osteogenic differentiation in the presence of PBLG/n-HA for BTE. The cell-biomaterial interactions were analyzed using cell proliferation, SEM and CMFDA dye extraction techniques. Osteogenic differentiation of ADSC was confirmed using alkaline phosphatase activity (ALP), mineralization (ARS) and dual immunofluorescent staining using both ADSC marker protein and Osteocalcin, which is a bone specific protein. The utmost significance of this study is the bioactive PBLG/n-HA biomolecule introduced on the polymeric nanofibers to regulate and improve specific biological functions like adhesion, proliferation and differentiation of ADSC into osteogenic lineage. This was evident from the immunostaining and CMFDA images of ADSCs showing cuboidal morphology, characteristic of osteogenic lineage. The observed results proved that the PLLA/PBLG/Col/n-HA scaffolds promoted greater osteogenic differentiation of ADSC as evident from the enzyme activity and mineralization profiles for bone tissue engineering.

PMID: 22048006 [PubMed - as supplied by publisher]


A combination of shear and dynamic compression leads to mechanically induced chondrogenesis of human mesenchymal stem cells.
Source:  Pub Med
Monday, 29 November 1999 17:00

A combination of shear and dynamic compression leads to mechanically induced chondrogenesis of human mesenchymal stem cells.

Eur Cell Mater. 2011;22:214-25

Authors: Schätti O, Grad S, Goldhahn J, Salzmann G, Li Z, Alini M, Stoddart MJ

Abstract
There is great interest in how bone marrow derived stem cells make fate decisions. Numerous studies have investigated the role of individual growth factors on mesenchymal stem cell differentiation, leading to protocols for cartilage, bone and adipose tissue. However, these protocols overlook the role of biomechanics on stem cell differentiation. There have been various studies that have applied mechanical stimulation to constructs containing mesenchymal stem cells, with varying degrees of success. One critical fate decision is that between cartilage and bone. Articular motion is a combination of compressive, tensile and shear deformations; therefore, one can presume that compression alone is unlikely to be a sufficient mechanical signal to generate a cartilage-like tissue in vitro. Within this study, we aimed to determine the role of shear on the fate of stem cell differentiation. Specifically, we investigated the potential enhancing effect of surface shear, superimposed on cyclic axial compression, on chondrogenic differentiation of human bone marrow-derived stem cells. Using a custom built loading device we applied compression, shear or a combination of both stimuli onto fibrin/polyurethane composites in which human mesenchymal stem cells were embedded, while no exogenous growth-factors were added to the culture medium. Both compression or shear alone was insufficient for the chondrogenic induction of human mesenchymal stem cells. However, the application of shear superimposed upon dynamic compression led to significant increases in chondrogenic gene expression. Histological analysis detected sulphated glycosaminoglycan and collagen II only in the compression and shear group. The results obtained may provide insight into post-operative care after cell therapy involving mesenchymal stromal cells.

PMID: 22048899 [PubMed - in process]