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目的探讨体外直接诱导HSF6人胚胎干细胞(humanem bryonic stem cells,hESCs)分化为神经细胞的方法。方法采用直接的方法,在1%血清培养条件下,顺序添加bFGF、RA和Forskolin,诱导HSF6人ESCs分化为神经细胞。结果细胞发生神经样形态学改变,免疫荧光细胞化学分析结果显示,分化细胞表达神经干细胞特异性标志分子——巢蛋白(nestin),以及神经元标志分子——β微管蛋白Ⅲ(neuron-specific class Ⅲ beta-tubulin,TuJ1)。实验组nestin阳性细胞数占(95.2±3.03)%,明显高于未添加诱导因子组的(31.6±4.93)%,差异有统计学意义(P<0.05)。结论本研究直接诱导hESCs分化为神经细胞,减少了常规经胚胎体(embryoid body,EB)的诱导方法而产生其它胚层细胞的机会,为进一步探索hESCs源性神经细胞的功能,以及为细胞替代治疗提供高纯度的hESCs源性神经细胞奠定了基础。
Objective To investigate the method of direct differentiation of human embryonic stem cell line HSF6 into neural cells in vitro. Methods Human embryonic stem cells (HSCs) were induced to differentiate into neurons by direct addition of bFGF, RA and Forskolin in 1% serum. Results The neuron-like morphological changes occurred in the cells. Immunofluorescent cytochemical analysis showed that the differentiated cells expressed neural stem-specific marker nestin and neuron-specific marker neuron-specific class III beta-tubulin, TuJ1). The number of nestin positive cells in the experimental group was (95.2 ± 3.03)%, which was significantly higher than that in the group without induction factor (31.6 ± 4.93)%, the difference was statistically significant (P <0.05). Conclusions This study directly induced the differentiation of hESCs into neurons, reduced the chances of inducing other germinal cells by inducing conventional embryoid bodies (EBs). In order to further explore the function of hESCs-derived neurons and the role of cell replacement therapy The basis for providing high purity hESCs-derived neural cells has been established.