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目的:研究大鼠骨髓间充质干细胞(MSCs)体外定向分化为多巴胺(DA)能神经元过程中细胞周期相关蛋白的表达变化。方法:体外分离、扩增和鉴定大鼠骨髓MSCs。碱性成纤维细胞生长因子(bFGF)预诱导24 h后,用1μmol/L全反式维甲酸(ATRA)和50 ng/ml GDNF联合诱导6 d,检测神经元特异性烯醇化酶(NSE)、胶质纤维酸性蛋白(GFAP)、酪氨酸羟化酶(TH)、增殖细胞核抗原(PCNA)、Bcl-2、细胞周期蛋白依赖性激酶(CDK2)、细胞周期蛋白(Cyclin)B1的表达。结果:诱导后大鼠骨髓MSCs能分化为具有典型神经元形态的细胞,大鼠MSCs经诱导后PCNA、CDK2、Cyclin B1的表达均较对照组明显减弱,Bcl-2表达与对照组无明显差别。结论:骨髓间充质干细胞体外开始分化后细胞增殖力下降。
OBJECTIVE: To study the changes of cell cycle related proteins during the differentiation of rat bone marrow mesenchymal stem cells (MSCs) into dopaminergic neurons in vitro. Methods: The bone marrow MSCs were isolated, expanded and identified in vitro. After induced by basic fibroblast growth factor (bFGF) for 24 h, neuron-specific enolase (NSE) was detected by the combination of 1 μmol / L all-trans retinoic acid (ATRA) and 50 ng / , Glial fibrillary acidic protein (GFAP), tyrosine hydroxylase (TH), proliferating cell nuclear antigen (PCNA), Bcl-2, cyclin dependent kinase (CDK2) and cyclin B1 . Results: MSCs could differentiate into cells with typical neuronal morphology after induction. The expression of PCNA, CDK2 and Cyclin B1 in rat MSCs was significantly weaker than that in control group after induction, and the expression of Bcl-2 was not significantly different from that in control group . CONCLUSION: BMSCs begin to differentiate and proliferate in vitro.