大鼠脑星形胶质细胞的体外培养及鉴定

Culture and identification of rat cerebral astrocyte in vitro

目的探讨新生大鼠大脑皮质星形胶质细胞的分离、培养以及鉴定方法。方法取出生1-2d的Wistar大鼠大脑皮质，用机械法和胰酶消化法分散细胞，制成细胞悬液，差速黏附处理后，将未贴附的细胞接种培养。待细胞铺满瓶底后，置摇床摇晃，舍弃含脱落细胞的培养液，细胞传代。GFAP免疫细胞化学染色鉴定。结果成功分离培养了原代星形胶质细胞，传代培养的星形细胞形态典型，纯度可达95％以上。结论成功建立了大鼠星形胶质细胞体外培养方法。

Objective To explore a method of isolation, culture and identification of astrocyte from cerebral cortex of neonatar rats. Methods The cerebral cortex was obtained from 2 to 3 days- old Wistal rat pups and prepared cell suspension by digestion with trypsin and mechanical digestion. After differential attachments, primary cells were cultivated to confluence. The cultures were shaken on an orbital shaker, forsook the supernatant and passaged. The passaged cells were immunocytochemieally stained with anti-GFAP antibody for identifying the astrocyte. Results The primary astrocyte was successfully cultured and identified. By passage,the purified astrocytes had the typical shapes with a purity up to 95%. Conclusion The method of culture in vitro of astrocyte is successfully established.