小鼠脑内星形胶质细胞体外培养方法的优化

目的： 优化小鼠脑内星形胶质细胞的体外培养方法，为星形胶质细胞的功能研究奠定实验基础。 方法： 取新生0～1 d的C57BL/6乳鼠大脑皮质，胰酶消化结合机械吹打方法，分离皮质细胞，制成细胞悬液并差速贴壁处理，待细胞接种24 h轻柔换液，以后每隔2～3 d换液，且每次换液前手动振荡洗涤2次，传代3次后，行胶质纤维酸性蛋白（glial fibrillary acidic protein,GFAP）免疫细胞化学染色及免疫荧光染色鉴定。 结果： 经过原代及传代培养，获得形态典型、纯度在95%以上的星形胶质细胞。结论： 优化后的体外培养星形胶质细胞方法，稳定有效，操作简便易行，并可获得高纯度的星形胶质细胞。

Optimization on the cultivation method in vitro of cerebral astrocytus from mice

Objective: To optimize the cultivation method of cerebral astrocytes from mice in vitro, in which develop an experimental basis for the functional study of astrocytus.  Methods: Cerebral cortex was obtained from 0 to 1d C57BL/6 mice and cell suspension was prepared by digestion with trypsin digestion and mechanical dissociation. After differential attachments, primary cells were cultivated for 24 h, so the fresh culture medium instead of the old was added softly. After then, the cell flask was washed shakely before the old culture medium was replaced with the fresh medium every two or three days. After three passages, astrocytus in the culture medium were stained with anti-glial fibrillary acidic protein (GFAP) antibody using immunocytochemical or immunofluorescence staining.  Results:  After primary cell culture and subculture, the purified astrocytes had the typical shapes with a purity beyond 95%.  Conclusion:  A reproducible and simple cultivation method in vitro of astrocytus was established, in which high purity astrocytes were obtained.