大鼠神经干细胞体外培养  

In vitro culture of rat neural stem cell

作　　者：曹翠丽[1] 阎蕴力[2] 杨春[3] 马常升[1] 石葛明[1] 郑力芬[2] 

机构地区：[1]河北医科大学基础医学研究所神经生物室 [2]河北医科大学基础医学研究所细胞生物室,050017 [3]河北医科大学基础医学研究所解剖教研室

出　　处：《脑与神经疾病杂志》2005年第3期165-168,共4页Journal of Brain and Nervous Diseases

基　　金：河北省教育厅资助课题:项目编号:2004113

摘　　要：目的:介绍一种较成熟的大鼠神经干细胞体外培养方法。方法:取新生大鼠(1d之内)侧脑室前角周围的脑组织,用EDTA:胰蛋白酶(1:1)消化,制成单细胞悬液,以2-5×105个细胞/ml接种于无血清培养基中。观察其增殖、分化情况,并用nestin、GFAP、NSE免疫细胞化学对细胞进行定性。结果:培养细胞呈悬浮生长,具有增殖能力,倍增时间为2d,nestin阳性。诱导分化后细胞贴壁生长,体积增大,并发出形态各异的突起。可见GFAP和NSE阳性细胞。结论:培养细胞经鉴定为神经干细胞。本方法简单、易操作,可行性强。Objective: To introduce a mature method for rat neural stem cell culture in vitro. Methods: Collecting the brain tissues around the anterior horn of lateral ventricle from newly born rats (within Id), then digested them with ED-TA and trypsin (1:1 mixed) for obtaining single cell suspension. The obtained cells were cultured in serum-free medium with 2-5×05/ml to investigate its proliferation and differentiation. The immunocytochemistry was performed to identify nestin, GFAP and NSE in the cells, respectively. Results: It was demonstrated that the cells grew in a manner of neu-rospheres and suspended in the medium. Those cells were shown to be nestin-positive, and had the proliferative ability which doubling time was 2 days. The cells became adherence growth with increasing size and stretching processes of different style, when they were induced to differentiation. There were GFAP-positive and NSE-positive cells among the differentiated cells. Conclusion: The cultured cells belong to neural stem cell and the introduced method is simple and reliable.

关 键 词：细胞体外培养 NESTIN 神经干细胞 体外培养方法 无血清培养基 免疫细胞化学 GFAP 培养细胞 侧脑室前角 单细胞悬液 新生大鼠 胰蛋白酶 EDTA 增殖能力 倍增时间 细胞贴壁 诱导分化 阳性细胞 NSE 脑组织 生长 

分 类 号：R135.12[医药卫生—劳动卫生] R743.31[医药卫生—公共卫生与预防医学]