大鼠海马神经干细胞的原代培养方法  

Primary Culture Method of Rat Hippocampal Neural Stem Cells

作　　者：丁海月 薛冰 刁华琼 魏丹 李小黎 DING Haiyue;XUE Bing;DIAO Huaqiong;WEI Dan;LI Xiaoli(Beijing University of Chinese Medicine Third Affiliated Hospital,Beijing 100029,China)

机构地区：[1]北京中医药大学第三附属医院,北京100029 [2]首都医科大学中心实验室

出　　处：《中西医结合心脑血管病杂志》2021年第6期949-951,共3页Chinese Journal of Integrative Medicine on Cardio-Cerebrovascular Disease

基　　金：国家自然科学基金面上项目(No.81673930)。

摘　　要：目的探索总结一种简单、高效的大鼠海马神经干细胞原代培养方法。方法选取新生24 h的SD大鼠,剥离出双侧海马,用机械吹打法制成细胞悬液,加入培养液,接种于培养皿中,3 d后半量换液,5~7 d后可传代。每天在荧光倒置显微镜下观察细胞的生长状况,采用免疫荧光法对第2代神经干细胞进行鉴定。结果本方法能准确分离出海马组织,培养出的神经干细胞中无其他混杂的细胞生长,经鉴定呈兔抗巢蛋白(Nestin)阳性和5-嗅-2-脱氧尿苷(BrdU)阳性。结论运用本方法获得的海马神经干细胞纯度高、细胞活力高、增殖能力强。Objective To explore and summarize a simple and efficient method for primary culture of rat hippocampal neural stem cells.Methods The 24 hour newborn SD rats were selected,the bilateral hippocampus was stripped,and the cell suspension was prepared by mechanical pipetting.The culture medium was added and inoculated into the petri dish.After 3 days,the medium was changed in half,and it could be passaged after 5 to 7 days.The growth of cells were observed under a fluorescent inverted microscope every day,and the second-generation neural stem cells were identified using immunofluorescence.Results This method can accurately isolate hippocampal tissue,and the cultured neural stem cells have no other mixed cell to growth.It is identified as Nestin positive and BrdU positive.Conclusion The hippocampal neural stem cells obtained by this method with high purity,high cell viability,and strong proliferation ability.

关 键 词：海马 神经干细胞 原代培养 实验研究 

分 类 号：Q813.11[生物学—生物工程]