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目的研究体外利用全反视黄酸(RA)诱导小鼠胚胎干细胞(mESC)分化为神经细胞的培养方法。方法悬滴3天转悬浮1天两者结合的培养方法得到胚体(EBs),再利用RA对其处理4天后,进行免疫组织化学、RT-PCR以及兴奋性功能的检测,观察神经细胞获得的效率及其生理特性。结果利用RA诱导方法得到的神经细胞,免疫组化表明nestin蛋白表达的阳性率为(53.49±6.02)%,且RT-PCR法分析其能表达nestin、glutaminase和Brn-3等神经细胞特异基因,同时在Glu刺激下具有与脑来源神经元相似的特性和功能。结论利用本实验诱导方法,获得神经细胞的比例较高,且具有与神经元相似的生物学特性。
Objective To study the culture method of alloanti-retinoic acid (RA) -induced mouse embryonic stem cell (mESC) differentiation into nerve cells in vitro. METHODS: Embryoid bodies (EBs) were obtained by a combination of two days after the suspension was suspended for 3 days. The cells were treated with RA for 4 days. Immunohistochemistry, RT-PCR and excitatory function were detected. The efficiency and its physiological characteristics. Results The positive rate of nestin protein expression was (53.49 ± 6.02)% by immunohistochemistry using RA-induced method. The expression of nestin, glutaminase and Brn-3 were detected by RT-PCR. At the same time, it has the same characteristics and functions as brain-derived neurons under the stimulation of Glu. Conclusion The proportion of neural cells obtained by the method of this experiment is high, and has the similar biological characteristics as neurons.