【摘要】 目的 探讨成年大鼠骨髓间充质干细胞（ａｄｕｌｔ ｒａｔ ｍａｒｒｏｗ ｍｅｓｅｎｃｈｙｍａｌ ｓｔｅｍ　ｃｅｌｌｓ，ｒＭＳＣｓ）的体外 增殖和特异性诱导分化为神经元样细胞的能力。方法 分别采用３种不同的诱导方法体外定向诱导第三至五代的 ｒＭＳＣｓ向神经元样细胞分化，并分别应用相差显微镜观察神经元样细胞和免疫细胞组化检测神经元样细胞所表达 的神经元特异性标志［神经元特异性烯醇化酶（ｎｅｕｒｏｎ ｓｐｅｃｉｆｉｃ ｅｎｏｌａｓｅ，ＮＳＥ）和神经丝蛋白（ｎｅｕｒｏｆｉｌａｍｅｎｔ，ＮＦ）］及 星形胶质细胞特异性标志［胶质纤维酸性蛋白（ｇｌｉａ ｆｉｂｅｒ ａｃｉｄ ｐｒｏｔｅｉｎ，ＧＦＡＰ）］，并进行神经元样细胞定量分析。结 果 所采用的３种定向诱导方法都能使ｒＭＳＣｓ特异性诱导分化为神经元样细胞，此神经元样细胞都能特异性地表 达出ＮＳＥ和ＮＦ，而不表达ＧＦＡＰ。经过定量计数分析发现用上述方法处理ｒＭＳＣｓ后出现ＮＳＥ阳性的细胞约为 ７５．５％±３．５％，出现ＮＦ阳性的细胞约为７７．２％±２．８％。结论ｒＭＳＣｓ能在体外扩增、传代，并能被定向诱导分 化为神经元样细胞。更多还原

【Abstract】 Objective To investigate whether the adult rat marrow mesenchymal stem cells (rMSCs) could be expanded and induced to differentiate into neuron-like cells in vitro or not. Methods Three different treatment protocol has been used to induce rMSCs differentiate into neuron-like cells in the present study. The first and sec- ond treatment protocol: rMSCs was pretreated with preinduction media consisting of IMEM/15%FCS/1mmol/L 2-mercaptoethanol (2-ME) or monothiglycerol for 24h,then was induced with induction media consisting of IMEM/ 5mmol/L2-ME or monothiglycerol for 5h, respectively. The third treatment protocol: rMSCs was pretreated with preinduction media consisting of IMEM/15%FCS/10ng/ml basic fibroblastic growth factor(bFGF) for 24h, then was induced with induction media consisting of IMEM /2% dimethvlsulfoxide (DMSO) /200Umol/L butvlated hy- droxyanisole(BHA) /25mmol/L Kcl /1μmol/ hydrocortisone /5μg/ml insulin for 5h to 6h. The differentiating cells was observed by phase-contrast microscopy and detected by immunocytochemistry. Results All three treatment protocols can induce rMSCs differentiate into neuron-like cells in the present study. The neuron-like cells expressed neuron-specific marker .such as neuron-specific enolase (NSE) and neurofilament (NF) . but could not express glia fiber acid protein(GFAP).which is astrocytes specific marker. With these differentiation protocol , about 75. 5± 3.5% and 77. 2%±2.8% of the rMSCs NSE positive and NF positive cells, respectively. Conclusions rMSCs could be explanded, passaged, and induced to differentiate into neuron-like cells in vitro.更多还原