大鼠脑皮质未成熟星形胶质细胞的分离、鉴定及原代培养  

Isolation, identification and primary culture of immature astrocyte of rat brain cortex

作　　者：吴炎宇 侯博[1] 王伯丞 郭英[1] 

机构地区：[1]中山大学附属第三医院神经外科,广州510630

出　　处：《中华临床医师杂志（电子版）》2015年第11期101-104,共4页Chinese Journal of Clinicians(Electronic Edition)

基　　金：广东省自然科学基金重点项目(S2012020010919);高等学校博士学科点专项科研基金(20120171110083);广东省产学研项目(2011B090400098)

摘　　要：目的探讨大鼠脑皮质未成熟星形胶质细胞的分离、鉴定及原代培养技术。方法采用差速贴壁法清除成纤维细胞及酶消化去除少突胶质细胞污染，应用免疫荧光进行细胞鉴定。取不同时间点的细胞在倒置显微镜下观察细胞形态变化，双重免疫荧光染色观察细胞生长过程的分化状态。结果细胞形态观察提示：6～14d细胞呈均匀扁平多角状，16d后部分细胞肥大增生，呈成熟状态。免疫组化染色显示：GFAP（＋）细胞达97％。免疫荧光双重标记发现：16d后细胞表达vimentin和nestin这两种蛋白减弱。结论本实验技术可获得纯度高，形态和功能良好，表达稳定的大鼠脑皮质未成熟星形胶细胞。Objective To investigate the isolation, identification and primary culture of immature astrocytes of rat cerebral cortex. Methods Method of differential adherence and trypsin digestion were used to remove fibroblasts and oligodendrocytes, and identified by immunofluorescence. Morphology were observed by inverted phase contrast microscope at different time points. Double immunofluorescenee staining was used in the observation of cells differentiation. Results Over 97% of the cells turned out to be GFAP positive. At day 6-14, cells was evenly, fiat, homed in morphology, and partially grew mature at day 16. Immunofluorescence double labeling showed a decrease of vimentin and nestin expression at day 16. Conclusion ＂The technology of our experiment is efficient to abtain high purity, morphology and function well, and stable expression immature astroeytes cells from rat brain cortex.

关 键 词：神经胶质原纤维酸性蛋白质 波形蛋白 巢蛋白 未成熟星形胶质细胞 

分 类 号：R329.2[医药卫生—人体解剖和组织胚胎学]