成人骨髓间质干细胞体外分化为神经元样细胞  

Human mesenchymal stem cells differentiate into neuron-like cells in vitro

作　　者：姚晓黎[1] 张成[1] 卢锡林[1] 张为西[1] 冯善伟[1] 柳太云[1] 

机构地区：[1]中山大学附属第一医院神经科,广州510080

出　　处：《广东医学》2004年第5期504-506,共3页Guangdong Medical Journal

基　　金：卫生部临床学科重点项目 (编号 :2 0 0 13 2 1) ;广东省科技计划项目 (编号 :ZKM0 5 5 0 1S)

摘　　要：目的　探讨成人骨髓间质干细胞 (hMSC)体外增殖和不同诱导剂对hMSC定向分化为神经元样细胞的作用。方法　体外分离hMSC ,培养扩增。分别用参芪液、β -巯基乙醇 (β -ME)和复合成分诱导剂体外定向诱导hMSC分化为神经元。于诱导后 1 ,3和 5h分别应用相差显微镜观察神经元样细胞和进行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)的表达 ,并进行神经元样细胞定量分析。结果　加入 3种诱导剂后 ,可使hMSC特异性的分化为神经元样细胞。免疫组化显示诱导出的神经元样细胞NSE ,NF阳性 ,GFAP阴性。神经分化的定量分析显示诱导后 1 ,3和 5h ,参芪液诱导NSE ,NFH阳性细胞分别为 5 0 3% ,6 3 5 % ,79 6 %和 4 6 5 % ,5 8 9% ,77 5 %。经 β -ME诱导NSE ,NFH阳性细胞分别为 4 4 6 % ,6 0 9% ,77 2 %和 4 3 7% ,5 6 1 % ,75 6 %。经复合成分诱导NSE ,NFH阳性细胞分别为 5 0 5 % ,6 2 3% ,80 5 %和 4 5 1 % ,5 9 1 % ,78 8%。结论　hMSC可在体外扩增、传代 。ObjectiveTo investigate whether the human mesenchymal stem cells(hMSC) could be expanded and induced into neuron-like cells with different inducers. MethodshMSCs were separated and expanded in culture medium. hMSCs were induced to differentiate with different inducers shenqiye, β-mercaptoethanol(β-ME) and complex inducers. After 1 h , 3 h and 5 h of induction, neuron-specific enolase(NSE), neurofilament(NF), and glial fibrillary acidic protein(GFAP) were detected by immunocytochemistry. The ratio of neuron-like cells was counted by phase-contrast microscopy. ResultshMSCs differentiated into neuron-like cells after being treated with inducers shenqiye, β-ME and complex inducers. hMSCs exhibited neuronal phenotype. The expression of NSE and NF in the neuron-like cells was positive, but the glial astrocyte marker GFAP did not express. After 1 h, 3 h and 5 h of induction with shenqiye, the ratio of NSE positive cells was 50 3%,63 5% and 79 6% respectively. NF positive rate was 46 5%, 58 9% and 77 5% respectively. After 1 h, 3 h and 5 h of induction with β-ME, the ratio of NSE positive cells was 44 6%,60 9% and 77 2% respectively. NF positive rate was 43 7%, 56 1% and 75 6% respectively. After 1 h, 3 h and 5 h of induction with complex inducers, the ratio of NSE positive cells was 50 5%,62 3%,80 5% and 45 1%,59 1%,78 8% respectively. ConclusionhMSC can be explanded, passaged and induced into neuron-like cells by different inducers.

关 键 词：成人 骨髓间质干细胞 体外 细胞分化 神经元样细胞 

分 类 号：R329.2[医药卫生—人体解剖和组织胚胎学]