Wnt3a影响神经干细胞分化的体外研究

目的探讨Wnt3a对胚胎大鼠海马神经干细胞（NSCs）体外分化的影响。方法采用机械分离、无血清传代培养法从胎鼠海马中获得NSCs，使用免疫荧光法对其干细胞特性及其受体Fzd3蛋白表达进行鉴定，观察Wnt3a对NSCs体外分化的影响。结果海马NSCs表达特异性标志物巢蛋白及胞膜蛋白Fzd3；体外诱导分化，Wnt3a处理组神经元及星形胶质细胞分化的比例分别为11．25％±0．62％和56．26％±4．82％，而对照组则为8．54％±0．48％和168．42％±5．54％；组间分化差异具有统计学意义（P〈0．05）。结论体外环境下Wnt3a能够促进NSCs向神经元分化，并抑制其向星形胶质细胞分化。

Effect of Wnt3a on differentiation of neural stem cells derived from hippocampal tissues of embryonic rats brains in vitro

Objective To investigate the effect of Wnt3a on the differentiation of neural stem cells(NSCs).Methods NSCs were separated mechanically from hippocampal tissues of embryonic rats brain and cultured in serum-free medium.The expression of nestin and transmembrane protein frizzled-3(Fzd3) in NSCs were detected by the immunofluorescence technique in order to identify NSCs.NSCs were divided into two groups,i.e.experiment group(NSCs were cultured in the medium containing Wnt3a 100 ng/ml) and control group(NSCs were cultured in the medium without wnt3a).NSCs differentiation was induced and observed.Results The NSCs expressing nestin and Fzd3 were successfully acquired.The percentages of β-tubulin Ⅲ positive cells(neurons) and glial fibrillary acidic protein-positive cells(astrocytes) were 11.25%±0.62% and 56.26%±4.82% respectively in the experimental group,and 8.54%±0.48% and 68.42%±5.54% respectively in the control group.There was significantly difference in the percentages of the neurons and astrocytes between both the groups(P <0.05).Conclusion The differentiation of NSCs into neurons may be promoted and into astrocytes inhibited by Wnt3a in vitro.