【摘要】 目的观察Nurr1基因修饰原代培养的神经干细胞(NSCs)体外诱导分化为多巴胺能神经元的作用。方法用电穿孔法将携带Nurr1基因的质粒转染P3代NSCs并进行潮霉素筛选,24h后免疫细胞化学检测转染效果;并对转染后的NSCs进行传代培养至P6代,检测Nurr1基因修饰的NSCsNurr1表达效果,并进行分化实验,分化培养7d后免疫细胞化学检测酪氨酸羟化酶(TH)的表达。对照组选用未转染的NSCs。结果电穿孔法转染的NSCs24h后免疫细胞化学检测到Nurr1高表达;传代至P6代,检测到Nurr1较高表达;对照组呈弱表达。分化实验表明:Nurr1基因修饰的NSCs分化的神经细胞中TH阳性比例占27.20%±7.12%,对照组为5.74%±1.81%;差异有显著性(P<0.01)。结论Nurr1基因修饰可以促进原代培养的NSCs向多巴胺能神经元方向分化。更多还原

【Abstract】 Objective To observe the effects of dopaminergic neurons differrentiatied from primary neural stem cells(NSCs) modified by Nurr1 gene cultured in vitro. Methods Transfected recombinating expression plasmid(pcDNA3.1-hygro-Nurr1) into P3 NSCs by the method of electroporation and used hygromycin bolting. Detected the effects of the transfection by the method of immuocytochemistry after 24 h. Passaged the transfected NSCs to P6 and detected the effects of the expression of Nurr1 in the NSCs modified by Nurr1. Detected the expression of Nurr1 by the method of immuocytochemistry after differential culture. Selected the nontransfected NSCs. Results High expression of Nurr1 in the NSCs transfected by electroporation was detected by the method of immuocytochemistry after 24 h. Deutohigh expression of Nurr1 was detected in passage 6. Poor expression of Nurr1 was detected in the control groups. Differential experiment indicates that, the percentage of tyrosine hydroxylase(TH) positive staining neurons which differentiated from NSCs modified by Nurr1 gene were 27.20%±7.12%, and 5.74%±1.81% for the control group respectively. The variance was significance(P<0.01)/ Conclusion Modify of Nurr1 gene could promote NSCs cultured primarily to differentiate into TH positive neurons.更多还原