体外诱导兔骨髓间充质干细胞定向分化为软骨细胞的实验研究

In vitro study of rabbit chondrocytes differentiation from bone marrow mesenchymal stem cells

目的:研究兔骨髓间充质干细胞(BMMSCs)向软骨细胞分化的潜能。方法:将兔髂骨骨髓液进行原代和传代培养,以高糖DMEM无血清特定培养液诱导(含TGF-β210ng/ml、地塞米松7mol/L-10mol/L、维生素C50μmol/L),相差显微镜下观察细胞形态变化,免疫组织化学染色检测软骨特异性II型胶原表达。结果:BMMSCs诱导后细胞体外扩增能力显著降低,细胞形态由成纤维样细胞梭形向多角形、多边形或类圆形转变,诱导21天后细胞形态变化最为显著,II型胶原免疫组化染色深而均匀。结论:在适当条件下,体外培养的BMMSCs可定向诱导分化为软骨细胞。

Objective：To explore the differentiation of rabbit bone marrow measenchymal stem ceils （BMMSCs） into chondrocytes like ceils in vitro. Methods： After primary culture and subcultures of BMMSCs, the cell were cultured in serum - free chondroinductive medium containing TGF - β2, dexamethasone, ascorbic acid. The growth of the cultured BMMSCs were observed by phase - microscope. Immunohistochemical staining was applied to detect the expression of collagen type II. Results ： The ceils changed from a spindle - like fibroblastic appearance to a polygonal shape, the capability of proliferation was down markedly. Immunohistochemical staining of collagen Ⅱ were positive for the passage, especially in the 21st day. Conclusion： The BMMSCs can differentiate into chondrocytes -like cells in vitro.