【摘要】 目的改进大鼠脑皮质星形胶质细胞体外培养方法,获得纯度较高的大鼠脑皮质星形胶质细胞。方法取出生后1d内SD大鼠大脑皮质,用机械吹打法使细胞分散制成初细胞悬液,10μm尼龙膜过滤除去成纤维细胞后接种。待7d细胞铺满瓶底后,"十"字形震荡培养液5min后传代接种。传代2次后行GFAP免疫组化染色鉴定。结果通过微孔尼龙膜过滤和传代前"十"字形震荡培养液等操作可以得到形态典型、纯度较高的星形胶质细胞。通过纯度计算可得离体培养的星形胶质细胞的纯度平均值为95.49%(n=6)。结论改进的星形胶质细胞体外培养方法,降低了实验操作难度,方法稳定有效,得到的星形胶质细胞纯度较高。更多还原

【Abstract】 Objective To develop an improved method for the cultivation of astrocytes from the cerebral cortex of SD rats.Methods Cerebral cortex was obtained from 1 day old rats.The cells were mechanically dissociated form the tissues and the cell suspension was pass through a nylon membrane(pore size:10 μm) to remove contaminating fibroblasts.Cells were seeded to plastic flasks and cultured for 7 days till subculture.Before trypsinization,the flasks were hand shaken for 5 minutes and then subculture.After 2 passages,astrocytes in the culture were stained with anti-GFAP antibody using immunocytochemical staining method.Results Using this method,the purity of astrocytes was 95.49%(n=6).Conclusion A simple and reproducible method was developed to produce astrocytes with high purity in the culture.更多还原