胎鼠脑神经干细胞的分离、培养和鉴定  

Isolation,cultivation and identification of neural stem cells in fetal rats

作　　者：刘磊[1] 刘恒方[1] 张敏[1] 郭亚培[1] 吴世陶[1] 高灵利[2] 

机构地区：[1]郑州大学第五附属医院神经内科,河南省450052 [2]河南大学第一附属医院神经内科

出　　处：《江苏医药》2015年第7期753-755,F0002,共4页Jiangsu Medical Journal

基　　金：河南省教育厅科学技术研究重点项目(12A320064)

摘　　要：目的研究胎鼠脑皮质神经干细胞(NSCs)的分离、培养及鉴定方法。方法从孕15d胎鼠的大脑皮层和海马区脑组织中获取NSCs,在含有B27、表皮生长因子(EGF)和碱性成纤维生长因子(bFGF)的DMEM/F12无血清培养液中培养;传代后用5%胎牛血清培养液诱导NSCs分化。结果体外分离培养的NSCs在无血清培养液中形成大量的神经球。经3-5代传代的细胞生长稳定。经巢蛋白染色鉴定,大部分为阳性细胞,神经细胞球经胎牛血清培养液贴壁培养后可分化为神经元特异烯醇化酶、胶质纤维酸性蛋白和半乳糖脑苷脂表达阳性的细胞。结论从孕15d胎鼠大脑皮质和海马组织分离出的NSCs具有自我更新能力和多向分化潜能,其在5%胎牛血清培养液中具有向神经元和神经胶质细胞分化的潜能。Objective To study the techniques for the isolation,cultivation and identification of neural stem cells（NSCs）in fetal rats.Methods The NSCs of the cerebral cortex and hippocampus were isolated from fetal rats with 15 days of gestational age.The NSCs were cultured in DMEM/F12serum-free culture medium containing B27,epidermal growth factor（EGF）and basic fibroblast growth factor（bFGF）,which then were added 5% fetal bovine serum to induce neural stem cells differentiation.Results The nerve balls were seen in a great number in serum-free medium,which were growing stably after passing 3-5generations.Identifyed by nestin dyeing,the majority of the cells were positive,which could express neuron specific enolization enzyme,glial fiber acidic protein and galactocerebroside.Conclusion The NSCs of the cerebral cortex and hippocampus from fetal rats with15 days of gestational age have the ability of self-renewal and multi-directional differentiation potential,which have a potential of differentiating into neurons and glial cells when induced by 5%fetal bovine serum.

关 键 词：神经干细胞 

分 类 号：R329[医药卫生—人体解剖和组织胚胎学]