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首页> 外文期刊>Cell and Tissue Research >Creation of an in vitro microenvironment to enhance human fetal synovium-derived stem cell chondrogenesis
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Creation of an in vitro microenvironment to enhance human fetal synovium-derived stem cell chondrogenesis

机译：建立体外微环境以增强人类胎儿滑膜衍生的干细胞软骨生成

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Our aim was to assess the feasibility of the sequential application of extracellular matrix (ECM) and low oxygen to enhance chondrogenesis in human fetal synovium-derived stem cells (hfSDSCs). Human fetal synovial fibroblasts (hfSFs) were characterized and found to include hfSDSCs, as evidenced by their multi-differentiation capacity and the surface phenotype markers typical of mesenchymal stem cells. Passage-7 hfSFs were plated on either conventional plastic flasks (P) or ECM deposited by hfSFs (E) for one passage. Passage-8 hfSFs were then reseeded for an additional passage on either P or E. The pellets from expanded hfSFs were incubated in a serum-free chondrogenic medium supplemented with 10 ng/ml transforming growth factor-β3 under either normoxia (21% O2; 21) or hypoxia (5% O2; 5) for 14 days. Pellets were collected for evaluation of the treatments (EE21, EE5, EP21, EP5, PE21, PE5, PP21, and PP5) on expanded hfSF chondrogenesis by using histology, immunostaining, biochemistry, and real-time polymerase chain reaction. Our data suggest that, compared with seeding on conventional plastic flasks, hfSFs expanded on ECM exhibit a lower expression of senescence-associated β-galactosidase and an enhanced level of stage-specific embryonic antigen-4. ECM-expanded hfSFs also show increased cell numbers and an enhanced chondrogenic potential. Low oxygen (5% O2) during pellet culture enhances hfSF chondrogenesis. Thus, we demonstrate, for the first time, the presence of stem cells in hfSFs, and that modulation of the in vitro microenvironment can enhance hfSDSC chondrogenesis. hfSDSCs might represent a promising cell source for cartilage tissue engineering and regeneration.
机译：我们的目的是评估顺序应用细胞外基质（ECM）和低氧来增强人胎儿滑膜衍生干细胞（hfSDSCs）软骨形成的可行性。人类胎儿滑膜成纤维细胞（hfSFs）的特征在于，发现其包括hfSDSCs，这由它们的多分化能力和间充质干细胞典型的表面表型标记所证明。将第7代hfSFs平板接种在hfSFs（E）沉积的常规塑料烧瓶（P）或ECM上一次。然后将第8代hfSFs播种到P或E上，以供其他传代。将来自扩增的hfSFs的沉淀物在常氧（21％O sub> 2 ; 21）或缺氧（5％O 2 ; 5）进行14天。通过使用组织学，免疫染色，生物化学和实时聚合酶链反应，收集药丸以评估对扩大的hfSF软骨形成的处理（EE21，EE5，EP21，EP5，PE21，PE5，PP21和PP5）。我们的数据表明，与在常规塑料瓶中播种相比，在ECM上扩增的hfSFs表现出较低的衰老相关β-半乳糖苷酶表达，并具有较高水平的阶段特异性胚胎抗原4。 ECM扩增的hfSFs还显示出细胞数量增加和软骨形成能力增强。颗粒培养过程中的低氧（5％O 2 ）增强hfSF软骨形成。因此，我们首次证明了hfSFs中存在干细胞，并且体外微环境的调节可以增强hfSDSC软骨形成。 hfSDSCs可能代表软骨组织工程和再生的有希望的细胞来源。

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