OLN-93细胞系分化条件探讨  

Study on the optimal differentiation culture media for OLN-93 cell culture

作　　者：周颖 苏敏 杰吉甫[2] 符辉[1] Zhou Ying;Su Min;Jie Jifu;Fu Hui(Department of Human Anatomy and Histo-Embryology,School of Basic Medical Sciences,Wuhan University,Wuhan 430071,China;Department of Anatomy,Health School of Bayinguoleng Mongolian Autonomous Prefecture,Korla 841000,China)

机构地区：[1]武汉大学基础医学院人体解剖和组胚胚胎学系,武汉430031 [2]新疆巴音郭楞蒙古自治州卫生学校解剖学系,库尔勒841000

出　　处：《中国组织化学与细胞化学杂志》2020年第2期125-130,共6页Chinese Journal of Histochemistry and Cytochemistry

基　　金：武汉市科技局应用基础研究计划(2017060201010168);国家级大学生创新创业项目(S2018301759)。

摘　　要：目的OLN-93细胞是实验室常用的少突胶质前体细胞细胞系,多被用于研究少突胶质细胞的分化。本文来探讨OLN-93细胞系的最佳分化条件。方法分别用DMEM、DMEM+三碘甲状腺原氨酸(T3)、DMEM+T3+N2、DMEM/F12、DMEM/F12+T3、DMEM/F12+T3+N2等6种不同培养基条件对OLN-93细胞系进行分化处理,于处理后的第3d和第6d观察细胞形态变化并通过免疫荧光技术鉴定细胞分化情况,同时通过检测髓鞘主要标志物MBP、CNP基因的mRNA和蛋白质的表达情况来分析细胞的分化状态,从而筛选出OLN-93细胞的最优分化培养基。结果比较OLN-93细胞在6种不同条件下的分化情况,免疫荧光、qPCR和Western Blot检测显示出基本一致的结果,DMEM培养基中的细胞分化状态最好,其次为DMEM+T3组,而DMEM+T3+N2和DMEM/F12+T3+N2组最差。结论DMEM培养基为适宜OLN-93细胞生长分化的最佳培养条件,其他成分如F12、N2均会不同程度的抑制细胞的分化。我们实验发现OLN-93已经丧失了对T3的反应,这一点在使用OLN-93细胞的实验研究中需要加以考虑。Objective To explore the optimization of the differentiation media for OLN-93,a common oligodendrocyte precursor cell line widely used to study the differentiation of oligodendrocytes in labs throughout the world.Methods OLN-93 cells were differentiated with 6 different media DMEM,DMEM+triiodothyronine(T3),DMEM+T3+N2,DMEM/F12,DMEM/F12+T3,and DMEM/F12+T3+N2.3 days and 6 days after treatment(D3 and D6)were used as 2 time points to observe the cell morphological changes and for further measurement.Immunofluorescent staining,qPCR and western blot techniques were used to detect myelin gene(MBP and CNP)expression to analyze the cell differentiation status.And thus the optimal differentiation medium for OLN-93 cells was determined.Results Comparing the differentiation status of OLN-93 cells under 6 different conditions,immunofluorescence,qPCR and Western Blot detection showed generally the same results.The best cell differentiation status of OLN-93 cell was obtained in DMEM medium,followed by DMEM+T3.While the cell differentiation status in DMEM+T3+N2 and DMEM/F12+T3+N2 groups was the worst.Conclusion DMEM medium is the best culture medium for OLN-93 growth and differentiation.Both F12 and N2 inhibit OLN-93 cell differentiation.OLN-93 cells have lost their responses to T3 differentiation signal.This fact should be considered in OLN-93 experiments.

关 键 词：OLN-93细胞 少突胶质细胞 分化 三碘甲状腺原氨酸 

分 类 号：R-331[医药卫生]