碱性成纤维细胞生长因子等体外诱导成人骨髓间充质干细胞分化为神经元样细胞的研究

Human mesenchymal stem cells differentiate into neuron-like cells with bFGF

目的 成人骨髓间充质干细胞 (h MSCs)体外定向诱导分化为神经元样细胞。方法 采用 Percoll分离液离心分离 h MSCs,体外扩增 ,分别采用含碱性成纤维细胞生长因子 (b FGF)和 2 -巯基乙醇 (2 - ME)等无血清DMEM诱导 h MSCs分化为神经元。免疫组化鉴定神经元烯醇化酶 (NSE)、神经丝蛋白 (NF)、胶质纤维酸性蛋白(GFAP)。结果 h MSCs在体外扩增传至 5代后 ,流式细胞仪显示 99.5 % ,97.8% ,98.8% h MSCs表面抗原 CD2 9、CD4 4、CD90 表达阳性。接种到 12孔板 ,3d后加入 b FGF和 2 - ME联合或 2 - ME单种诱导剂诱导后 ,h MSCs胞体收缩 ,突起伸出 ;免疫组化显示诱导出的神经元样细胞 NSE、 NF表达阳性 ,GFAP阴性。

Objective To investigate the differentiation from human mesenchymal stem cells(hMSCs)into neuron like cells. Methods hMSCs were separated from rib marrow with Percoll reagent and expanded in culture medium. hMSCs were induced to differentiate into neurons with basic fibroblast growth factor (bFGF) and 2 mercaptoethanol(2 ME),respectively.Neuron specific enolase(NSE),neurofilament(NF),nestin,glial fibrillary acidic protein(GFAP) were detected by immunohistochemistry.Results hMSCs were expanded to be undifferentiated cells in culture for more than 5 passages.Flow cytometry showed that 99.5%,97.8%,98.8% hMSCs expressed strongly antigens of CD 29 ,CD 44 ,CD 90 which was identical to human bone marrow derived MSCs.Immunohistochemistry staining showed that the induced different passage hMSCs expressed NSE,NF and nestin,but did not express GFAP.Conclusion It suggests that hMSCs can be differentiated into neurons in vitro.