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目的观察血管内皮生长因子(vascular endothelial growth factor,VEGF)在人胚胎干细胞分化为神经元过程中是否发挥作用及相关机制。方法人胚胎干细胞经拟胚体向神经元分化,分为3组:A组为常规诱导组,B组为常规诱导+VEGF(10ng/mL)作用组,C组为常规诱导+VEGF(10ng/mL)+VEGFR2/Fc嵌合体(10ng/mL)作用组;用RT-PCR、免疫荧光法检测各阶段细胞标志物,计算并用流式细胞仪检测各组不同阶段细胞阳性率。结果用RT-PCR分别检测到OCT4、Nestin、MAP2表达;免疫荧光法检测显示B组产生神经干细胞、分化为神经元的阳性率明显高于A、C两组,差异有统计学意义(P<0.01),A、C两组之间无显著性差异(P>0.05);流式细胞仪检测与免疫荧光法相似。结论人胚胎干细胞体外分化过程中血管内皮生长因子通过血管内皮生长因子受体2来促进神经干细胞增殖及向神经元分化。
Objective To observe whether vascular endothelial growth factor (VEGF) plays a role in the process of human embryonic stem cells differentiating into neurons and related mechanisms. Methods Human embryonic stem cells (NSCs) differentiated into neurons by embryoid bodies were divided into 3 groups: group A was a conventional induction group, group B was induced by conventional induction of VEGF (10ng / mL), group C was conventional induction of VEGF (10ng / mL) + VEGFR2 / Fc chimera (10ng / mL). The cell markers were detected by RT-PCR and immunofluorescence. The positive rate of cells in different stages of each group was calculated and analyzed by flow cytometry. Results The expression of OCT4, Nestin and MAP2 were detected by RT-PCR. The immunofluorescence assay showed that the positive rate of neural stem cells differentiated into neurons in group B was significantly higher than that in groups A and C (P < 0.01). There was no significant difference between A and C groups (P> 0.05). Flow cytometry was similar to immunofluorescence. Conclusion Vascular endothelial growth factor (VEGF) promotes the proliferation of neural stem cells and differentiate into neurons through the vascular endothelial growth factor receptor 2 during the differentiation of human embryonic stem cells in vitro.