胎鼠神经干细胞分离培养和鉴定  

Culture and identification of neural stem cells from embryonic rats in vitro

作　　者：田志红[1] 黄国伟[1] 姬长珍[1] 张绪梅[1] 王广雷[1] 张文治[2] 苏心[2] 

机构地区：[1]天津医科大学公共卫生学院营养与食品卫生教研室,天津300070 [2]天津市神经外科研究所

出　　处：《天津医科大学学报》2007年第4期476-478,共3页Journal of Tianjin Medical University

基　　金：国家自然科学基金资助项目(30571563)

摘　　要：目的:建立胎鼠神经干细胞体外培养方法,观察神经干细胞生长特点。方法:采用含碱性成纤维细胞生长因子(bFGF)和表皮生长因子(EGF)的无血清培养基对大鼠胚胎大脑组织细胞悬液进行培养,细胞免疫荧光方法对神经干细胞鉴定,MTT法测定不同细胞密度的OD值,绘制生长曲线。结果:从胎鼠脑组织分离培养的细胞巢蛋白阳性。MTT结果显示细胞接种密度以2(×104～105)/ml生长良好(r=0.95,P<0.05)。结论:无血清培养基分离培养的胎鼠脑组织神经干细胞具有自我更新和增殖能力,巢蛋白细胞免疫鉴定为神经干细胞。Objective： To establish the methods of isolation, culturing of neural stem cells（NSCs）in vitro and observe the characteristics of NSCs growth. Methods： NSCs were isolated and cultured by microdissection, mechanical blowing and serum-free suspension culture. Fluorescence immuncytochemistry was applied to identify the NSCs. MTr method was used to dedect the growth of NSCs of three different density. Results：Nestin-positive cells could be found from the cell of embryonic rat brain. MTr results showed that NSCs of density of 2×（10^4-10^5）/ml grew well （r=0.95, P〈0.05）. Conclusion：The method can be used to isolate and culture NSCs from embryonic rat brain, the cell identified as NSCs possess the ability of proliferation and self-reproduction.

关 键 词：神经干细胞 细胞培养 免疫荧光 胎鼠 

分 类 号：Q813.11[生物学—生物工程]