单只小鼠来源的大脑皮层星形胶质细胞的体外原代培养

摘要

Objective: To establish a system of single mouse-based culture of the primary cerebral astrocytes.Methods: A single mouse of different genotypes within postnatal 4 days was used to obtain the bilateral cortex in aseptic condition.The cells were then mechanically dispersed into suspension and fibroblasts were removed by the method of differential adhesion.Nine days after being confluent, the cells were passaged and stained with GFAP antibody for astrocytes.Results: The astrocyte was successfully isolated.The proportion of astrocyte was more than 95％, and the number of cells was more than 106.Conclusion: The improved method can easily establish a system to culture astrocytes in vitro from only one newborn mouse.%目的:建立单只小鼠来源的星形胶质细胞体外原代培养体系.方法:取单只新生小鼠,无菌操作下取双侧大脑皮质,彻底剥除脑膜,制备细胞悬液,差速贴壁法去除成纤维细胞,悬液原代培养9d左右,待细胞融合成单层并铺满瓶底后,消化传代,胶质纤维酸性蛋白免疫组织化学显色鉴定星形胶质细胞.结果:分离培养出的星形胶质细胞生长良好,纯化率达95%以上,细胞数目能达到106以上.结论:采用该改进的方法能简捷地用单只小鼠大脑皮层建立星形胶质细胞的体外培养及研究体系.