体外直接诱导人胚胎干细胞向神经细胞分化的研究

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。

In vitro direct neural cells differentiation of human embryonic stem cells

Objective To explore the in vitro direct neural cell differentiation of human embryonic stem cells （hESCs）. Methods We developed a protocol that allowed direct neural differentiation of hESCs in 1% serum culture system. A stepwise addition of factors, as bFGF, retinoic acid （RA） and then Forskolin （FN） was employed for the conversion. Results Most of differentiated cells showed neural properties in cell morphology. Immunocytochemistry revealed most of the differentiated cells expressed neural stem cell marker-nestin, and neuron marker-neuron-specific class Ⅲ beta-tubulin （TuJ1）. Furthermore, compared to the control group （31.6±4.93）%, the nestin positive cells in experimental group was up to （95.2±3.03）%（P0.05）. Conclusions We introduced an efficient induction system for directly in vitro neural differentiation of hESCs,so as to avoid unwanted cells from other germ layers via embryoid body formation.This will benefit to further functional analysis of differentiated neural cells and to cell replacement therapies with hESCs.