SD大鼠星形胶质细胞的原代培养

Primary cultivation of SD rat cerebral astrocyte

目的设计原代培养SD大鼠星形胶质细胞。方法 取新生第2～3d SD红皮鼠,无菌操作下取大脑皮质,剪碎、胰酶消化结合机械吹打使细胞分散,网筛过滤,差速粘附处理去除成纤维细胞,将未粘附的细胞悬液接种培养9～12d,置摇床,舍弃含脱落细胞的细胞悬液,以去除少突胶质细胞和小胶质细胞。细胞传代,GFAP免疫组化染色鉴定。结果 成功分离培养了原代神经胶质细胞,并进行了星形胶质细胞的纯化,GFAP鉴定星形胶质细胞比例为95％以上。结论 建立星形胶质细胞体外培养方法,总结操作关键,以此为实验材料对于比较学研究是非常有帮助的。

Objective Astrocyte-enriched culture from postnatal SD rat cerebral tissue was prepared. Methods Cerebral cortex was dissected and trypsinized as well as mechanical method to make an initial cell suspension from 2 to 3 day-old SD rat pups. Use differential attachment to reduce the amount of contaminating fibroblasts. After an initial culture period between 10 and 14 day, the cultures were shaken on an orbital shaker, to reduce oligodendrocytes and microglial cells. Passaged cells were immunocytochemically stained with anti-GFAP antibody to identify astrocytes. Results We successfully obtained the primary astrocyte culture. The preparations appeared >95 pure. Conclusion The method is established and the preparations should significantly aid in comparative study.