成人骨髓源性神经干细胞中nestin表达的实时定量RT-PCR检测  

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

作　　者：朱汝森[1] 徐如祥[2] 姜晓丹[2] 蔡颖谦[2] 邹雨汐[2] 杜谋选[2] 

机构地区：[1]广东医学院附属医院神经外科,广东湛江524001 [2]南方医科大学珠江医院神经外科,广东广州510282

出　　处：《海南医学》2009年第8期1-4,共4页Hainan Medical Journal

基　　金：国家自然科学基金(U0632008)

摘　　要：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 a-dult 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2.04×102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。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 invitre, 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 flasks. hMSCs could be efficiently differentiated into Md - NSCs by induction culture invitro. Md - NSCs did not adhere to the surface of tissue culture flasks, and the cell clones form neurospherelike structures. Nestin was found to be over- expressed in Md- NSCs compared with that in hMSCs, and the magnitude of the increased expression was 2.04 ×10^2. Conclusion Md - NSCs is not aggregates of hMSCs and have very different biological characteristics from hM- SCs.

关 键 词：神经干细胞 神经巢蛋白 基因表达 实时定量RT—PCR 

分 类 号：R741[医药卫生—神经病学与精神病学]