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

Bioprocess Research and Assay Development, Tengion Inc., Winston-Salem, North Carolina 27103, USA.
Tissue Engineering Part C Methods (Impact Factor: 4.64). 05/2011; 17(8):843-60. DOI: 10.1089/ten.tec.2010.0697
Source: PubMed


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.

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    • "These studies underline that SVF may be a promising adjuvant population boosting the efficacy of conventional AFT. However, there seems to be a lack of standardization in the technique due to poorly controlled factors that may introduce biases, such as different devices to isolate SVF, sub-standardized adherence between SVF and AFT, their number and the same heterogeneous nature of SVF [6, 12]. "
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    ABSTRACT: Autologous fat transfer (AFT) is a procedure for adipose tissue (AT) repair after trauma, burns, post-tumor resections and lipodystrophies still negatively impacted by the lack of graft persistence. The reasons behind this poor outcome are unclear and seem to involve damages in either harvested/transplanted mature adipocytes or on their mesenchymal progenitors, namely adipose stromal/stem cells (ASC), and due to post-transplant AT apoptosis and involution. A rabbit subcutaneous AT regeneration model was here developed to first evaluate graft quality at different times after implant focusing on related parameters, such as necrosis and vasculogenesis. Standard AFT was compared with a strategy where purified autologous ASC, combined with hyaluronic acid (HA), assisted AFT. Five million of autologous ex vivo isolated CD29+, CD90+, CD49e+ ASC, loaded into HA, enriched 1 ml of AT generating an early significant protective effect in reducing AFT necrosis and increasing vasculogenesis with a preservation of transplanted AT architecture. This beneficial impact of ASC assisted AFT was then confirmed at three months with a robust lipopreservation and no signs of cellular transformation. By a novel ASC assisted AFT approach we ensure a reduction in early cell death favoring an enduring graft performance possibly for a more stable benefit in patients. Electronic supplementary material The online version of this article (doi:10.1007/s10495-013-0878-7) contains supplementary material, which is available to authorized users.
    Apoptosis 07/2013; 18(10). DOI:10.1007/s10495-013-0878-7 · 3.69 Impact Factor
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    • "Adipose is recognized as an endocrine organ with significant metabolic bioactivity. Adipose tissue is composed of adipocytes, vascular endothelial cells, pericytes, fibroblasts, macrophages, stem cells and progenitors with MSC-like bioactivity and smooth muscle-like cells [1-4]. Of these, MSC-like and smooth muscle-like cell populations are currently under active development for application in tissue engineering and regenerative medicine [5]. "
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    Lipids in Health and Disease 09/2011; 10:171. DOI:10.1186/1476-511X-10-171 · 2.22 Impact Factor
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    ABSTRACT: Mesenchymal stem cells (MSCs) show great promise for use in a variety of cell-based therapies. Because isolated primary mesenchymal stem cells are low in numbers, in vitro expansion is necessary. However, the expansion potential is limited and in vitro aging leads to loss of multipotency and replicative senescence. Stress induced by culture conditions is likely to be a major cause of replicative senescence and reduced multipotency of MSC and optimization of culture conditions might be able to reduce this. Caloric restriction (CR) is the only established method to delay aging and extend lifespan. In vitro caloric restriction experiments are rare, but have demonstrated beneficial effects. Therefore, we investigated the effect of culture medium glucose concentration on the proliferative and differentiation potential of mesenchymal stem cells. Reduction in glucose concentrations led to decreased apoptosis and an increased rate of MSC proliferation and increased the number and size of fibroblastic colonies in the colony-forming unit assay.
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