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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。
OBJECTIVE: To observe the natural degeneration time of rat knee chondrocytes cultured in vitro and to provide suitable target cells for the study of osteoarthritis. Methods: Chondrocytes were cultured, subcultured, passaged by toluidine blue and type Ⅱ collagen in chondrocytes of rats. Chondrocytes were characterized by Western Blot. The phenotype of chondrocytes was identified by Western Blot. Results: Compared with the first 4 passages, the expression of type Ⅱ collagen in the 5th generation of chondrocytes in the knee joints of rats decreased significantly (P <0.05). Conclusion: The normal chondrocyte phenotype was maintained in the first 4 passages of rat knee chondrocytes cultured in vitro , The fifth generation began degeneration, the first 4 generations can be used as chondrocytes target cells.