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目的探讨人脐带间充质干细胞(human umbilical cord mesenchymal stem cells,HuMSCs)向成肌细胞分化。方法贴壁法分离HuMSCs;流式细胞仪分析其表面抗原;透射电镜及原子力电镜观察细胞超微结构;5-氮杂胞苷诱导细胞向成肌细胞分化;逆转录聚合酶链式反应(RT-PCR)检测成肌细胞特异基因结蛋白(desmin)和重链肌球蛋白(myosin heavy chain,MHC)表达;蛋白质印迹法(Western Blot)检测特异蛋白desmin和MHC表达。结果贴壁法稳定从脐带分离出细胞;HuMSCs不表达CD31、CD34、CD45,强表达CD29、CD90、CD44、CD105,极低表达HLA-DR、CD40;5-氮杂胞苷诱导后干细胞表达成肌细胞特异基因及蛋白明显高于对照组。结论贴壁法分离HuMSCs可靠、纯度高;5-氮杂胞苷可诱导HuMSCs向成肌细胞分化。
Objective To investigate the differentiation of human umbilical cord mesenchymal stem cells (HuMMSCs) into myoblasts. Methods HuMSCs were isolated by adherence method. The surface antigens were analyzed by flow cytometry. The ultrastructures of the cells were observed by transmission electron microscopy and atomic force microscope. 5-azacytidine induced differentiation of myoblasts into myoblasts. Reverse transcriptase-polymerase chain reaction -PCR) were used to detect the expression of desmin and myosin heavy chain (MHC). The expression of desmin and MHC was detected by Western Blot. Results The adherence method was used to isolate cells from umbilical cord. HuMSCs did not express CD31, CD34 and CD45, but strongly expressed CD29, CD90, CD44 and CD105, and expressed HLA-DR and CD40. Muscle cell-specific genes and proteins were significantly higher than the control group. Conclusion Adherent method for the separation of HuMSCs is reliable and high purity. 5-Azacytidine can induce HuMSCs to differentiate into myoblasts.