成人骨髓源性神经干细胞中nestin表达的实时定量RT—PCR检测

目的建立快速获取成人骨髓源性神经干细胞（Human adult bone marrow—derived neural stem cells，Md—NSCs）的方法，并对Md—NSCs中神经巢蛋白nestin的表达进行定量比较分析。方法抽取成人志愿者全骨髓，梯度离心分离成人骨髓基质细胞（human adult bone marrow stromal cells，hMSCs），贴壁培养法纯化及扩增hMSCs；以神经干细胞培养基体外诱导培养hMSCs成为成人骨髓源性神经干细胞（Human adult bone marrow—derived neural stem cells，Md—NSCs）；以实时定量RT—PCR检测Md—NSCs中nestin基因的表达，以hMSCs作为对照组。结果hMSCs呈长梭形，贴壁生长，于体外诱导培养可快速有效地获取Md—NSCs，Md—NSCs不贴壁生长，由神经干细胞克隆组成神经球样结构；Md—NSCs中nestin基因的表达比hMSCs增高2．04×10^2倍。结论hMSCs经诱导分化成为Md—NSCs后细胞的生物特征发生显著改变，Md—NSCs高度表达神经巢蛋白nestin具有神经干细胞的特征，并非hMSCs的简单聚集成球。

Quantitative real-time RT-PCR analysis of nestin expression in human adult bone marrow-derived neural stem cells.

Objective To set up efficient method of generating human adult bone marrow-derived neural stem cells (Md-NSCs),and quantitatively test the nestin expression of Md-NSCs. Methods Human adult marrow stromal cells(hMSCs) were obtained from adult normal volunteers and differentiated into Md-NSCs invitro,and mRNA levels of nestin in Md-NSCs was investigated by quantitative real-time RT-PCR and was compared with that of hMSCs. Results hMSCs had a fusiformlike shape and adhered to the surface of tissue culture flas...