大鼠脑皮质星形胶质细胞体外培养方法的改进

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。

An improved method for the in vitro cultivation of astrocytes from cerebral cortex of SD rats

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.