纹状体组织提取液诱导人胚胎神经干细胞向多巴胺能神经元分化的实验研究

Effect of striatal extracts on inducing the NSC differentiate into dopaminergic neurons:an experimental study

目的体外培养人胚胎神经干细胞并诱导其向多巴胺能神经元分化。方法从临床引产的人胚胎（胎龄8~16周）海马组织中分离、培养人胚胎神经干细胞,对其进行诱导分化。诱导剂采用来源于20周胎龄胚胎的纹状体的组织提取液。实验分为2组：对照组无诱导剂,培养基为撤除生长因子的基础培养基;实验组培养基中加入纹状体组织提取液50μL/mL;于诱导分化的第7天终止诱导分化,采用标记TH的免疫荧光染色方法检测TH阳性细胞量,用RT-PCR方法检测TH-mRNA的表达情况。结果抗TH的免疫荧光染色结果为：对照组与实验组的TH阳性细胞率分别为（0.53±0.17）%和（7.38±0.84）%,两组相比差异有统计学意义（P〈0.05）。RT-PCR结果显示：对照组TH-mRNA表达不明显,实验组明显表达TH-mRNA,实验组与对照组的TH-mRNA表达差异亦有统计学意义（P〈0.05）。结论纹状体组织提取液具有促进人胚胎神经干细胞向多巴胺能神经元分化的作用。

Objective To culture human fetal neural stem cells in vitro, and then induce them to differentiate into dopaminergic neurons. Methods Hmnan fetal neural stem cells derived from the hippocampus were purified and induced by hmnan fetal striatal extracts. The induced differentiational rate of dopaminergic neurons was detected by immunocytoflurescence and the expression of THmRNA was determined by RT-PCR. Results The rate of TH ＋ cells of the experimental group was higher than that of the control group, and the expression of TH-mRNA was not obvious in the control group but was strong in the induced group. There were significant differences between the two groups. Conclusion Human fetal striatal extracts can effectively induce human fetal NSCs to differentiate into dopaminergic neurons in vitro.