大鼠胚胎神经干细胞与永生化神经干细胞系C17.2定向分化为神经元潜能的差异  

Variability of directed differentiation potential into neurons between neural stem cells from embryonic rats and immortalized neural stem cell line C17.2

作　　者：解俊领[1] 张玥[1] 黄小杰[1] 臧奕[2] 

机构地区：[1]华东师范大学生命科学学院,上海200062 [2]中国科学院上海药物研究所国家新药筛选中心,上海201203

出　　处：《解剖学报》2012年第5期581-587,共7页Acta Anatomica Sinica

基　　金：国家自然科学基金资助项目(81173033)

摘　　要：目的对比大鼠早期胚胎神经干细胞(NSCs)与永生化神经干细胞系C17.2(C17.2-NSC)定向分化为神经元潜能的差异,确立适用于诱导NSCs定向分化为神经元高内涵药物筛选的NSCs筛选模型。方法C17.2-NSC、17d胚胎海马NSCs(E17-NSC)及11d胚胎大脑皮层NSCs(E11-NSC)均设定对照组和实验组,对照组与实验组的细胞在含有2%(v/v)B27的DMEM/F12培养液中,分别经0μmol/L和1μmol/L维甲酸(RA)在37℃、5%CO2常规培养条件下诱导分化5d,通过免疫组织化学技术检测对照组与实验组中NSCs或前体细胞特异性标志蛋白巢蛋白(Nestin)及神经元特异性标志蛋白βⅢ微管蛋白(Tuj1)表达量的差异。结果与对照组相比,C17.2-NSC经1μmol/L RA诱导后未定向分化为神经元,而E17-NSC及E11-NSC经1μmol/L RA诱导后可有效定向分化为神经元。结论相对于C17.2-NSC,大鼠早期胚胎NSCs定向分化为神经元的潜能更强,适用于诱导NSCs定向分化为神经元的高内涵药物筛选。Objective To compare directed differentiation potential into neurons between neural stem cells（NSCs） from early embryonic rats and immortalized neural stem cell line C17.2（C17.2-NSC） , and to establish a screening assay on the level of NSCs for high content screening in order to discover drugs that induce directed neurons differentiation of NSCs. Methods Three types of cells, the C17.2-NSC, 17 days embryonic hippocampus NSCs and 11 days embryonic cerebral cortex NSCs were used in this study. Cells from each type were divided into the control and experimental groups, and induced to differentiate by 0 μmol/L and 1 p, mol/L retinoic acid （RA） for 5 days under DMEM/F12 medium containing 2% （V/V） B27 and 37℃ , 5% CO2 normal culture condition. The variability of expression level of nestin and βⅢ tubulin （Tujl） , which are the specific markers of neural stem and progenitor cells and neurons, respectively, in both control group and experimental group were detected through immunohistochemistry. Results Compared with the control group, C17.2-NSC did not differentiate into neurons under the induction of 1 p, mol/L RA, but 17-day embryonic hippocampus NSCs and l 1-day embryonic cerebral cortex NSCs effectively differentiated into neurons under the same treatment. Conclusion The directed differentiation capacity of early embryonic NSCs into neurons is higher than that of C17.2-NSC, thus early embryonic NSCs are suitable for high content drug screening which aims to discover new types of compounds that induce directed neurons differentiation of NSCs.

关 键 词：神经干细胞 神经元 细胞分化 维甲酸 巢蛋白 BIU微管蛋白 免疫组织化学 大鼠 

分 类 号：R338[医药卫生—人体生理学]