来源于人胚室下区神经干/祖细胞的贴壁法培养和鉴定  

Culture and identification of neural stem/progenitor cells from human embryonic subependymal zone by adherent method

作　　者：姜晓荣[1] 王洪美[1,2] 陈琳[1,2] 田国忠[1] 包建玲[1] 任玉水[1] 黄红云[1,2] 郗海涛[1] 

机构地区：[1]北京市虹天济神经科学研究院 [2]北京康复中心神经外科,北京100144

出　　处：《解剖科学进展》2010年第4期307-310,共4页Progress of Anatomical Sciences

基　　金：国家自然科学基金资助项目(No30971527)

摘　　要：目的建立有效的人胚胎神经干/祖细胞分离及纯化方法以达到临床需要。方法无菌取胎脑室管膜下区组织,经反复机械吹打制备细胞混悬液,采用贴壁法进行体外培养、传代。传代后用Nestin进行细胞鉴定。结果应用贴壁法进行胎脑室管膜下区神经干/祖细胞的培养能够稳定传代20代以上;共聚焦显微镜下可见Nestin阳性细胞表达呈递增趋势,传至P3(passage3)阳性率为35%,P7为79%,P12为90%,P15为99%。传到前七代时分别做细胞存活率鉴定,可达(82.57±1.38)%。结论成功建立了来源于人胚室下区的神经干/祖细胞的贴壁法培养,为该细胞移植治疗神经系统疾病提供了技术方法支持。Objective To explore a practical method for separation and purification of human embryonic neural stem/progenitor cells to satisfy the clinical application. Methods Under aseptic condition, tissue from subependymal zone of fetal brain was taken, cell suspension was prepared after repeated mechanical percussion, then cells were cultured and passaged by adherent method in vitro and immunostained by nestin. Results The neural stem/progenitor cells could be passed stably for more than 20 generations. The number of nestin-positive cells increased progressively under confocal microscopy. The rates of the nestin positive cells were 35%（P3）, 79%（P7）, 90%（P12）, 99% （P15） respectively. The survival rate of the first seven-generations was （82.57±1.38）%. Conclusion The adherent culture method is a successful method to obtain neural stem / progenitor cells from human embryonic subependymal zone.

关 键 词：神经干/祖细胞 细胞培养 人胚胎 免疫细胞化学 

分 类 号：R321[医药卫生—人体解剖和组织胚胎学]