新生SD大鼠少突胶质细胞前体细胞的培养与分化

Culture and differentiation of rat oligodendrocyte precursor cells

目的 体外培养高纯度的少突胶质前体细胞（oligodendrocyte progenitor cells,OPCs）以及分化成熟的少突胶质细胞（Oligodendrocytes,OLs）.方法新生1 -2 d Sprague-Dawley（SD）大鼠的大脑皮层胶质细胞混合培养9d后采用恒温摇床振荡分离与差异贴壁方法并结合条件培养基纯化培养细胞.光学显微镜观察细胞形态;纯化培养3 d后免疫荧光染色鉴定细胞类型.结果 获得高纯度的少突胶质前体细胞以及成熟的少突胶质细胞,少突胶质前体细胞免疫荧光NG2＋A2B5双标阳性,成熟少突胶质细胞免疫荧光MBP染色阳性.结论 采用恒温摇床振荡分离与差异贴壁法并结合条件培养基可以得到高纯度的少突胶质前体细胞以及成熟少突胶质细胞.

Objective To develop a simple and efficient oligodendrocyte progenitor cells （OPCs） culture method that can produce a high yield of pure OLs. Methods Mixed glial cells of cortices from neonatal rats （1 to 2 d after born） were primarily cultured for 9 d, and cells were purified by horizontal orbital shaking, differ ential adhesion and defined culture media. Cell was observed by optical microscope. After three days, cells were identified by immumofluorescence staining with NG2, A2B5 and DAPI. Results Large quantities of highly purified OPCs （specific marker NG2and A2B5） and OLs （specific marker MBP） generate by a simple method. Conclusions The method of horizontal orbital shaking, differential adhesion and defined culture media is effective to obtain highly purified OPCs.