视神经星状胶质细胞的培养  

The culture of optic astrocytes

作　　者：顾蓓[1] 刘玉琴[1] 张永鹏[1] 张宏[1] 王春景[1] 苏小玲[1] 董继红[1] 

机构地区：[1]中国医学科学院基础医学研究所北京协和医学院基础学院,北京100005

出　　处：《中国当代医药》2009年第10期33-35,共3页China Modern Medicine

摘　　要：目的:探索建立视神经星状胶质细胞培养的最佳方法,观察该细胞体外水解酶表达情况。方法:分别取不同年龄WISTAR大鼠(出生4 d、7 d、成年)的视神经,利用胰蛋白酶加EDTA、胶原酶、木瓜酶、胰蛋白酶加胶原酶等不同方法消化,分别接种在0.2%明胶、鼠尾胶、多聚赖氨酸包被的培养瓶中,进行原代培养,观察细胞生长状况。利用免疫组化鉴定分离培养的细胞。连续传代观察其衰老情况。利用水解空斑法观察其水解酶表达。结果:几种方法均可见细胞生长,年龄4、7 d大鼠视神经要比成年大鼠视神经胶质细胞培养成功率高,利用胰蛋白酶与胶原酶混合消化要比其他消化方法细胞损失小,速度快,细胞成活率高。培养细胞呈典型星状胶质细胞形态,GAFP呈阳性。常规3 d传代一次,可传代40次。胶质细胞在蛋白膜上可形成明显的水解空斑。结论:利用胰蛋白酶与胶原酶混合消化法,从新生鼠取材,可成功培养视神经星状胶质细胞,体外可传代40代。该细胞在体外可分泌蛋白水解酶。Obejctive：AIM Finding out the optimal approach to culture the optic astrocytes and to eatablish the cell line. Methods：Optic nerves from different ages＇ Wistar rats（4 d, 7 d or adult） was isolated and digested with trypsin/EDTA, collagenase, papain or trypsirdcollagenase. Flasks pre-coated with gelatin, rat tail collagen or poly-lysine were employed to seed the cells respectively. The grown cells were characterized by GAFP immuno-histochemistry. Cells were continously passaged until senescence. Results：Cells could grow under all conditions. The optic astrocytes from new born rats （4 or 7 d） were more cuceesfully cultivated than those from adult rats. The application of both trypsin and eollagenase were more rapid with less cell loss and higher cell survival than any other method. The cultivated cells displayed specific morphology of astrocytes. And GAFP was positve. These cells could be cultivated through 40 passages in vitro. Conclusion：Using combination of trypsin and collagenenase for digestion and taking tissue from new born hosts optic nerve astrocytes could be successfully cultivated. The astrocytes secrete proteolytic enzymes in vitro.

关 键 词：星状胶质细胞 GAFP 大鼠 

分 类 号：R329.24[医药卫生—人体解剖和组织胚胎学]