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首页> 美国卫生研究院文献>Stem Cells Translational Medicine >Effects of Medium Supplements on Proliferation Differentiation Potential and In Vitro Expansion of Mesenchymal Stem Cells
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Effects of Medium Supplements on Proliferation Differentiation Potential and In Vitro Expansion of Mesenchymal Stem Cells

机译：补充培养基对间充质干细胞增殖分化潜能和体外扩增的影响

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Mesenchymal stem cells (MSCs) possess great potential for use in regenerative medicine. However, their clinical application may be limited by the ability to expand their cell numbers in vitro while maintaining their differential potentials and stem cell properties. Thus the aim of this study was to test the effect of a range of medium supplements on MSC self-renewal and differentiation potential. Cells were cultured until confluent and subcultured continuously until reaching senescence. Medium supplementation with fibroblast growth factor (FGF)-2, platelet-derived growth factor (PDGF)-BB, ascorbic acid (AA), and epidermal growth factor (EGF) both increased proliferation rate and markedly increased number of cell doublings before reaching senescence, with a greater than 1,000-fold increase in total cell numbers for AA, FGF-2, and PDGF-BB compared with control cultures. Long-term culture was associated with loss of osteogenic/adipocytic differentiation potential, particularly with FGF-2 supplementation but also with AA, EGF, and PDGF-BB. In addition FGF-2 resulted in reduction in expression of CD146 and alkaline phosphatase, but this was partially reversible on removal of the supplement. Cells expressed surface markers including CD146, CD105, CD44, CD90, and CD71 by flow cytometry throughout, and expression of these putative stem cell markers persisted even after loss of differentiation potentials. Overall, medium supplementation with FGF-2, AA, EGF, and PDGF-BB greatly enhanced the total in vitro expansion capacity of MSC cultures, although differentiation potentials were lost prior to reaching senescence. Loss of differentiation potential was not reflected by changes in stem cell surface marker expression.
机译：间充质干细胞（MSCs）具有用于再生医学的巨大潜力。但是，它们的临床应用可能会受到体外扩展其细胞数量同时保持其差异电位和干细胞特性的能力的限制。因此，本研究的目的是测试一系列补充培养基对MSC自我更新和分化潜能的影响。培养细胞直至融合，并连续传代培养直至衰老。补充成纤维细胞生长因子（FGF）-2，血小板衍生生长因子（PDGF）-BB，抗坏血酸（AA）和表皮生长因子（EGF）的培养基均能增加增殖速率，并显着增加衰老前细胞倍增的数量与对照培养相比，AA，FGF-2和PDGF-BB的总细胞数增加了1000倍以上。长期培养与成骨/脂肪细胞分化潜能的丧失有关，尤其是补充FGF-2以及AA，EGF和PDGF-BB。此外，FGF-2导致CD146和碱性磷酸酶的表达降低，但这在去除补充剂后可部分逆转。细胞始终通过流式细胞术表达包括CD146，CD105，CD44，CD90和CD71在内的表面标记，并且即使在失去分化潜能后，这些推定的干细胞标记仍持续表达。总体而言，添加FGF-2，AA，EGF和PDGF-BB的培养基大大增强了MSC培养物的总体外扩增能力，尽管在达到衰老之前已经失去了分化潜能。干细胞表面标志物表达的变化未反映分化潜能的丧失。

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