School of Chemical and Biological Engineering, Seoul National University, Gwanak-gu, Republic of Korea.
Biotechnol Appl Biochem. 2011 Jul-Aug;58(4):271-6. doi: 10.1002/bab.39.
This study was designed to evaluate the additive effects of transforming growth factor-beta3 (TGF-β3) and hyaluronic acid (HA) on chondrogenic differentiation of human mesenchymal stem cells (hMSCs). The hMSCs were cultured on collagen type I-, HA-, or fibronectin-coated cell culture dishes with or without TGF-β3 added to the culture medium. Four weeks after cell culture, chondrogenic differentiation of hMSCs was determined by evaluating the expression of cartilage-specific markers using real-time polymerase chain reaction, immunocytochemistry, and Western blot analysis. hMSCs cultured on HA-coated dishes with TGF-β3 supplementation revealed a prominent increase in collagen type II, aggrecan, and Sox9. When hMSCs were cultured without TGF-β3 supplementation, only hMSCs cultured on HA-coated dishes showed prominent expression of the cartilage-specific markers. This study shows that chondrogenic differentiation of hMSCs can be enhanced additively by interactions with both a specific cell-adhesion matrix and a soluble growth factor.
本研究旨在评估转化生长因子-β3(TGF-β3)和透明质酸(HA)对人骨髓间充质干细胞(hMSCs)软骨分化的协同作用。hMSCs 在胶原 I 型、HA 或纤维连接蛋白包被的细胞培养皿上培养,培养基中添加或不添加 TGF-β3。细胞培养 4 周后,通过实时聚合酶链反应、免疫细胞化学和 Western blot 分析评估软骨特异性标志物的表达来确定 hMSCs 的软骨分化。在添加 TGF-β3 的 HA 包被培养皿上培养的 hMSCs 中,II 型胶原、聚集蛋白聚糖和 Sox9 的表达明显增加。当 hMSCs 培养中不添加 TGF-β3 时,仅在 HA 包被培养皿上培养的 hMSCs 表现出明显的软骨特异性标志物表达。本研究表明,hMSCs 的软骨分化可以通过与特定的细胞黏附基质和可溶性生长因子的相互作用得到协同增强。