新生大鼠大脑皮层少突胶质前体细胞的培养分离及鉴定

[目的] 探讨新生(sprague dawley,SD)大鼠大脑皮层少突胶质前体细胞在体外条件下的生长情况及培养分离,以获得纯化的少突胶质前体细胞,为今后细胞移植治疗研究提供良好的种子细胞.[方法] 出生48 h SD大鼠取大脑皮层进行混合胶质细胞的体外培养,原代培养第9～10 d时采用水平振荡、差速贴壁的方法分离、纯化少突胶质前体细胞,继续用定向培养基传代培养.相差显微镜观察少突胶质前体细胞在体外条件下的生长情况,免疫细胞化学技术进行细胞鉴定.[结果] 混合胶质细胞原代培养第9～10 d时出现明显的细胞分层,少突胶质前体细胞散布于单层的星形胶质细胞表面,胞体呈椭圆形或圆形,具有典型的双极突起,少数呈三极突起.经振荡分离后少突胶质前体细胞纯度达95%,少突胶质细胞特异性标记Olig02反应阳性,胶质纤维酸性蛋白抗体反应阴性.[结论] 新生SD大鼠大脑皮层原代培养中少突胶质前体细胞能够维持在未成熟阶段,细胞分层后通过振荡法及差速贴壁法可以获得较高纯度的少突胶质前体细胞.

Culture and isolation of oligodendrocyte precursor cells from neonatal rat cerebral cortices

[Objective]To observe the growth pattern of oligodendrocyte precursor cells(OPCs) in the primary culture from neonatal rat cerebral cortices and study the methods of cell culture and isolation in order to obtain purified OPCs for the experiments of cell transplantation.[Method]The mixed glial cells from the cerebral cortices of 48-hour-old Sprague-Dawley(SD) rats were cultured in vitro.Until 9-10 days in vitro OPCs were isolated and purified by the shaking process and differential adhesion,and then continue...