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人诱导性多能干细胞定向分化为神经干细胞的实验研究

Differentiation of induced pluripotent stem cells into neural stem cells

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【作者】 冯年花谢安娄远蕾阮琼芳郭菲汪泱

【Author】 FENG Nian-hua1,2,XIE An1,LOU Yuan-lei1,RUAN Qiong-fang1,GUO Fei1,WANG Yang1 (1.Institute of Urology,Nanchang University;2.Medical Department,Graduate School of Nanchang University,Nanchang 330006,China)

【机构】 南昌大学泌尿外科研究所南昌大学研究生院医学部

【摘要】 目的观察人诱导性多能干细胞(iPS细胞)定向分化为神经干细胞(NSCs)的潜能。方法用维甲酸(RA)诱导人iPS细胞向NSCs分化。倒置显微镜下观察iPS细胞的形态变化,RT-PCR检测NANOG、OCT4、SOX2的表达;免疫组织化学方法检测Nestin、SOX2、β-TubulinШ和GFAP的表达。结果 RA诱导后第4天,贴壁的拟胚体出现了早期神经祖细胞特有的神经管样结构并不断增多,细胞表达神经巢蛋白Nestin,而对照组未观察到神经管样结构。神经管样结构内的细胞能分化为β-TubulinШ阳性的神经元,但GFAP阳性的星形胶质细胞少见。结论人iPS细胞具有定向分化为NSCs的潜能,并能模拟神经发育过程。

【Abstract】 Objective To observe the differentiation potential of induced pluripotent stem(iPS) cells to neural stem cells(NSCs).Methods The differentiation process was carried out through retinoic acid(RA) inducing procedure.Cell transformation was observed using inverted microscope.RT-PCR was used to measure the expression of pluripotent genes.Specific markers of neural linage cells such as Nestin,SOX2,β-Tubulin Ш,GFAP were also detected by immunostainning.Results Neural tube-like constructions appeared 4 days after retinoic acid(RA) induction,these cells expressed Nestin and could differentiate to β-Tubulin Ш positive neurons and few GFAP positive astrocytes.There was no neural tube-like constructions in control group.Conclusion Human iPS cells can differentiate to neural stem cells and imitate the development process of nervous system.

【基金】 国家自然科学基金(30960385);江西省自然科学基金(2007GZY1470)
  • 【文献出处】 基础医学与临床 ,Basic & Clinical Medicine , 编辑部邮箱 ,2010年12期
  • 【分类号】R329
  • 【被引频次】7
  • 【下载频次】625
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