成年大鼠侧脑室下区星形胶质细胞的培养与纯化

Culture and purification of adult rat subventricular zone astrocytes in vitro

目的:探讨SD大鼠侧脑室下区星形胶质细胞(AST)的体外培养与纯化方法。方法:将差速贴壁后SD大鼠侧脑室下区AST置于含有B27添加剂的DMEM/F12中培养,采用低浓度血清(2.5～5ml/L)调整法对AST进行纯化。观察AST生长变化,采用形态学和GFAP免疫细胞化学染色进行细胞和纯度鉴定。结果:体外培养的大鼠侧脑室下区星形胶质细胞呈纤维状聚集或类圆形散在生长,GFAP染色呈阳性反应,细胞胞浆着色较深,高倍视野下细胞骨架形态清晰可见,细胞核不着色。在体外培养14～15d时可以获得91%的AST。结论:差速贴壁结合低浓度血清调整法是一种简单实用的AST体外培养纯化方法。

Objective：To explore a simple and pragmatic method to obtain sufficient astrocytes（ASTs） from subventricular zone by using different attachment rates in harvesting cells with the combination of serum limited methods.Methods：SVZs from subventricular zone of rats were cultured with DMEM/F12 culture solution containing 2% B27 and further purified by adjusting serum at low concentration（2.5%～5%） in combination with serum limited methods.The growth conditions of SVZs were observed.And then GFAP immunocytochemical staining was used to estimate the purity of the cells.Results：Primary cultured SVZs showed positive for GFAP immunocytochemical staining.They appeared to be fibroblast-shaped and oval in vitro.The purity of SVZs reached-91% on 14～15 d of culture.Conclusion：The method of different attachment rates in combination with serum limitation is a simple and effective to culture and purify SVZs.