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目的在体外建立稳定有效的软骨细胞分化模型。方法复苏ATDC5细胞,在倒置显微镜下观察细胞形态及其生长情况,细胞90%融合时分别用不含Vc和含Vc的诱导培养基培养进行分化诱导。诱导21 d,阿力新蓝染色,RT-PCR检测Ⅱ、Ⅹ型胶原表达进行鉴定。结果含50μg/mL Vc的诱导培养基诱导1周便可见明显的软骨小结,细胞产生软骨基质明显增多,Ⅱ及Ⅹ型胶原表达明显增高,且Ⅹ型胶原表达呈提前。结论利用ATDC5细胞系可成功建立软骨细胞分化体外模型。
Objective To establish a stable and effective chondrocyte differentiation model in vitro. Methods ATDC5 cells were resuscitated and the morphology and growth of ATDC5 cells were observed under an inverted microscope. Differentiation induction was induced by induction medium containing Vc and Vc at 90% confluency. Induced for 21 days, stained with alitake blue, and detected by RT-PCR for the expression of collagen type X and type X collagen. Results After induced by 50μg / mL Vc for 1 week, obvious cartilage nodules were observed. The cartilage matrix was significantly increased, the expressions of type Ⅱ and type Ⅹ collagen were significantly increased, and the expression of type Ⅹ collagen was premature. Conclusion The in vitro model of chondrocyte differentiation can be established successfully using ATDC5 cell line.