人胚胎纹状体组织提取液对人胚胎神经干细胞分化方向的影响

目的对人胚胎神经干细胞(NSCs)进行分离、培养,研究纹状体组织提取液对神经干细胞分化方向的影响。方法从临床因故引产的人胚胎(胎龄8~16周)海马组织中分离、培养人胚胎神经干细胞,将其分离纯化并进行抗Nestin染色鉴定后分为两组进行诱导分化实验。A组:以基础培养基作为对照组;B组:基础培养基 人胚胎纹状体组织提取液(50ml/L)。分化的第7d取出细胞爬片,分别进行抗神经特异性烯醇化酶(NSE)免疫细胞化学染色,抗5-羟色氨(TH)免疫荧光化学染色和RT-PCR方法检测TH-mRNA的表达。结果培养的海马细胞经抗Nestin染色后呈Nestin阳性,证明为神经干细胞。诱导分化后对照组与实验组的NSE阳性细胞率分别为(21.89±2.17)%和(23.50±1.60)%,两组比较结果无统计学差异(P>0.05)。TH阳性细胞率分别为(0.53±0.17)%和(7.38±0.84)%,两组比较有统计学差异(P<0.05)。RT-PCR检测结果表明,TH-mRNA在对照组中无明显表达,纹状体组织提取液组神经干细胞分化后,TH-mRNA表达明显,两组比较有统计学差异(P<0.05)。结论在体外培养中,人胚胎纹状体组织提取液对神经干细胞向神经元分化无明显促进作用,但能促进人胚胎神经干细胞向多巴胺能神经元分化。

EFFECT OF HUMAN STRIATAL EXTRACTS ON NSCs DIFFERENTIATION

Objective To isolate and cultivate human embryo neural stem cells and study the effect of striatal extracts on inducing NSCs to differentiate. Methods\ The human embryo neural stem cells derived from hippocampus were isolated, purified and then induced by human embryo striatal extracts. The ratio of neurons and dopaminergic neurons were respectively detected by immunochemistry method and immunoflurescence, and the expression of TH-mRNA were evaluated by the method of RT-PCR. Results\ Although the NSE+ cells rate in the experimental group(23.50±1.60)% was higher than that in the control group(21.89±2.17)%, there was no significant statistical difference between them(P>0.05). The ratio of TH+ cells in the experimental group(7.38±0.84)% was higher than that in the control group(0.53±0.17)%,and there was significant statistical difference between the two group(P<0.05). The expression of TH-mRNA was not evident in the control group, but was positive in the experimental group. There was a significant difference between the two groups. Conclusion\ Human embryo striatal extracts cannot induce human embryo NSCs to differentiating into more neurons but it helps in