大鼠膝关节软骨细胞体外培养去分化时间的实验研究

目的:观察大鼠膝关节软骨细胞体外培养后自然退变时间,为骨关节炎研究提供合适靶细胞。方法:大鼠膝关节软骨细胞体外培养,传代,行甲苯胺蓝及Ⅱ型胶原染色鉴定软骨细胞,通过Western Blot检测其特征性Ⅱ型胶原的表达情况对传代软骨细胞表型进行鉴定。结果:大鼠膝关节第5代软骨细胞和前4代比较,Ⅱ型胶原表达明显减少(P<0.05),结论:大鼠膝关节软骨细胞体外培养前4代能维持正常的软骨细胞表型,第5代开始发生退变,前4代可以做为软骨细胞研究的靶细胞。

Degeneration Time of Chondrocytes Cultured in Vitro Derived from Knee Joint of Rats

Objective: To investigate the degeneration time of chondrocytes cultured in vitro derived from knee joint of rats in order to offer appropriate cells for the study on osteoarthritis.Methods: The chondrocytes derived from knee joint of rats were cultured and subcultured in vitro.Then the cultured cells were identified with toluidine blue staining and type Ⅱ collagen immunohistochemistry assay.The phenotype of chondrocytes were determined by assaying Type Ⅱ collagen with Western blot.Results: The synthesis of Type Ⅱ collagen as well as the cell proliferation rate descended notably in chondrocytes of genneration Ⅴ as compared with those of the first four generations(P<0.05).Conclusion: The degeneration of chondrocytes cultured in vitro derived from knee joint of rats were occurred after generation Ⅳ.The phenotype of chondrocytes from generationⅠto Ⅳcould sustain,and these chondrocytes could be served as suitable target cells to study ostearthritis.