大鼠脑皮质星形胶质细胞的纯化培养

目的获取纯度较高的大鼠脑皮质星形胶质细胞，用于进一步实验。方法取新生大鼠大脑，用酶消化结合机械吹打方法分离皮质细胞制成细胞悬液，差速贴壁法去除成纤维细胞，培养融合时，恒温摇床振荡法去除少突胶质细胞、小胶质细胞等，纯化后，观察星形胶质细胞生长变化规律，并通过GFAP免疫细胞化学方法鉴定星形胶质细胞。结果大鼠脑皮质细胞接种后24h全部贴壁，部分细胞已伸出突起；3～5d时细胞增殖最明显，细胞数目增加，体积增大，突起相互交织成网状；原代培养7～10d时细胞基本融合并开始分层生长。振荡纯化处理后，GFAP免疫细胞化学染色阳性细胞可达98％。结论用差速贴壁和恒温振荡方法可获得高纯度皮质源性星形胶席细胸．

The purification and culture of astrocytes from cerebral cortex of rats

Objective:To obtain purified astrocytes from rat cerebral cortex for further experiments.Methods:The neonatal rat brain was taken and the cortex was made into suspension with aids of enzyme-digestion and mechanical dissociation,and then reduced fibroblasts with differential attachment,oligodentrocytes and microglial cells with orbital shaker.After purification,the growth features of the cultured cells were observed and the astrocytes were identified with GFAP immunoreactivity.Results:Twenty-four hours after implantation,all cultured neonatal cerebral cortical cells adhered and some of them with small protrusions already.The cultured cells proliferated the fastest on days 3 through 5,when the cell number and volume increased rapidly,and their processes interlaced as a network.The cultured cells fused and got demix on days 7 through 10.Treated with orbital shaker and after passaged,the positive-GFAP immunoreactivity cells were counted no less than 98%.Conclusion:With differential attachment and orbital shaker in constant temperature,we successfully obtained the high purity astrocytes from neonatal rat cerebral cortex.