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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 the potential differences of directional differentiation of neural stem cells (NSCs) and immortalized neural stem cell line C17.2 (C17.2-NSC) into neurons in rats, and to establish a suitable screening method for inducing directional differentiation of NSCs into neurons NSCs screening model. Methods C17.2-NSC, 17d embryonic hippocampal NSCs (E17-NSC) and 11d embryonic cerebral cortex NSCs (E11-NSC) were set in the control group and the experimental group, the control group and the experimental group cells containing 2% (v / v) B27 were cultured in DMEM / F12 culture medium and differentiated with 0μmol / L and 1μmol / L retinoic acid (RA) respectively under the condition of 37 ℃ and 5% CO2 routine culture for 5 days. The control group and experiment were detected by immunohistochemistry The differences of the expression of Nestin and Tuj1 between NSCs or precursor cell-specific markers in the group. Results Compared with the control group, C17.2-NSC did not differentiate into neurons after 1μmol / L RA induction, while E17-NSC and E11-NSC induced neurotransmission after induced by 1μmol / L RA. CONCLUSIONS: Compared with C17.2-NSC, the potential of NSCs differentiated into neurons in early embryos is more suitable for the screening of high-content drugs that induce the directional differentiation of NSCs into neurons.