少突胶质前体细胞的培养及细胞缺氧缺糖模型的建立  

Oligodendrocyte precursor cells culture and oligodendrocyte oxygen/glucose deprived model establishment

作　　者：钟琳[1] 唐军[1] 姚裕家[1] 

机构地区：[1]四川大学华西第二医院儿科,成都610041

出　　处：《中国新生儿科杂志》2007年第4期218-220,I0003,共4页Chinese Journal of Neonatology

基　　金：教育部博士点基金项目资助(20050610071)

摘　　要：目的获取高纯度的大鼠少突胶质前体细胞系并建立细胞缺氧缺糖模型。方法采用振荡分离纯化法和化学限定无血清培养基在体外获取纯化的大鼠少突胶质前体细胞,免疫荧光法对各分化阶段特异的表面抗原A2B5、O4、O1进行细胞鉴定。利用无糖培养基和连二亚硫酸钠造成O4阳性的少突胶质前体细胞的缺氧缺糖损伤,观察细胞的形态学改变,MTT法测细胞存活率。结果获得纯度95%以上的大鼠少突胶质前体细胞,O4阳性少突胶质前体细胞出现缺氧缺糖损伤的形态学改变,并随缺氧缺糖时间延长而加剧;细胞存活率随缺氧缺糖时间的延长逐渐下降,30、60min和90min细胞存活率均低于正常对照组,差异有统计学意义(P<0.05)。结论振荡分离纯化法是体外培养大鼠少突胶质前体细胞的可靠方法,利用无糖培养基和连二亚硫酸钠可以建立细胞缺氧缺糖模型。Objective To obtain highly purified oligodendrocyte precursor cell lines in vitro and establish oxygen/glucose deprived （OGD） model of O4 positive oligodendrocyte. Methods The oligodendrocyte precursor cells were separated from astrocytes by orbital shaker, further purified by differential adhesion and cultured in chemically defined serum-free medium. Immunofluorescence assay was used to identify the separated cells. The OGD model of O4 positive oligodendrocyte was established by culturing cells with sodium dithionite in DMEM without glucose. Results The percentage of cultured oligodendrocyte precursor cells was more than 95%. The O4 positive oligodendrocyte cultured with sodium dithionite in DMEM without glucose presented with morphologic characteristics of OGD injury. And survival rate of the cells decreased gradually to lower levels than control at 30 min, 60 rain and 90 min （ P 〈 0.05 ） . Conclusions It is a reliable method to obtain oligodendrocyte precursor cells by separating and purifying with shaking and differential adhesion and culturing in chemically defined medium. The OGD model of O4 positive ohgodendrocyte can be estabhshed by cuhuring cells with sodium dithionite in DMEM without glucose.

关 键 词：少突神经胶质 细胞培养技术 模型 动物 

分 类 号：R-332[医药卫生]