大鼠大脑皮层星形胶质细胞体外培养方法改良

目的建立大鼠大脑皮层星形胶质细胞(As)的培养方法,以进一步对大脑皮层As进行体外实验研究。方法在M cCarthy方法基础上改用出生后5 d的SD大鼠的大脑皮层,制备单细胞悬液后,接种于培养瓶,培养箱中1 h后换瓶,3 d后换液,细胞融合成单层后传代,传3代后采用间接免疫荧光法监测As特异性标志胶质纤维酸性蛋白(GFAP)。阳性细胞确认为As。结果体外培养的大脑皮层As经历了贴壁、去除成纤维细胞、纯化和传3代,GFAP阳性细胞达95%以上。结论大脑皮层As体外培养成功,并有较高的纯度,为进一步进行As的研究创造了条件。

Improvement on culture in vitro of rat cerebral cortical astrocytes

Objective To establish a culture method of rat cerebral cortical astrocyte for further research on astrocyte in vitro.Methods Cerebral cortex of postnatal 5 days of SD rats were used basing on McCarthy method.Unicell suspension were prepared and inoculated in culture flask.Changed the culture flask after 1 hour and substratum was replaced after 3 days.Passage was done when cells syncretized to monolayer.Glial fibrillary acidic protein of the specifical sign of astrocyte was monitored by indirect immunofluorescence after three times passage.The masccline cells of glial fibrillary acidic protein were affirmed to be astrocyte.Results All cerebral cortical astrocytes cells which were cultured in vitro underwent adherence,fibroblast removed,depuration and passage three times,the rate of masccline cells of glial fibrillary acidic protein was above 95%.Conclusion Culture of cerebral cortical astrocytes in vitro is successful and there is higher purity.The condition was provided for further research of astrocytes.