基础医学与临床 ›› 2014, Vol. 34 ›› Issue (5): 579-582.

• 研究论文 •    下一篇

将人的皮肤成纤维细胞直接诱导为神经干细胞

贾伟丽1,李默1,吴剑宇1,王淑艳2,陈志国1,张愚2   

  1. 1. 首都医科大学宣武医院
    2. 首都医科大学宣武医院细胞室
  • 收稿日期:2013-12-26 修回日期:2014-01-07 出版日期:2014-05-05 发布日期:2014-04-28
  • 通讯作者: 张愚 E-mail:zhangyualex1975@tom.com
  • 基金资助:
    国家自然科学基金;国家重点基础研究发展计划973

Generation of induced neural stem/progenitor cells from human fibroblasts

  • Received:2013-12-26 Revised:2014-01-07 Online:2014-05-05 Published:2014-04-28
  • Contact: Y.Alex Zhang E-mail:zhangyualex1975@tom.com

摘要: 目的 探讨将人的皮肤成纤维细胞诱导为神经干细胞。方法 将成人的皮肤成纤维细胞用带有Pax6报告基因和绿色荧光蛋白的慢病毒感染,并经历其病毒载体的抗性基因潮霉素筛选后的细胞作为初始细胞,转入在维持神经干细胞生长发育、分化、保持功能稳定性和可塑性中起非常重要作用的10个基因Sox2、Klf4、c-Myc、Tcf3、Ascl1、Brn2、Neurog2、Foxg1、Hes1和Id1,对经过重新编程表达GFP绿色荧光蛋白的细胞用流式细胞仪进行筛选,并对其进行免疫荧光染色和分化的鉴定。结果 转入基因后成功诱导为神经干细胞(iNSCs)。iNSCs能表达神经干细胞的标志性基因(Nestin),能分化为神经元(β Ⅲ- tubulin)和星形胶质细胞(GFAP)。结论 通过转入外源因子的方式可以将人的皮肤成纤维细胞诱导为神经干细胞。

关键词: 关键词:诱导神经干细胞, 人的皮肤成纤维细胞, 重编程, 转录因子, 报告基因

Abstract: Objective To investigate induced neural stem/progenitor cells (iNSCs) from human fibroblasts. Methods Human fibroblasts were infected with lenti-virus expressing green fluorescent protein (GFP) under Pax6 promoter, followed by a hygromycin-resistant gene. Hygromycin-selected fibroblasts were used as starter cells and infected with virus encoding 10 transcription factors Sox2, Klf4, c-Myc, Tcf3, Ascl1, Brn2, Neurog2, Foxg1, Hes1, Id1, which play critical roles in the development, differentiation, functional stability, and plasticity of neural cells. The reprogrammed cells that were GFP-positive were sorted by using flow cytometer and characterized by fluorescence staining and differentiation assay. Results iNSC expressed NSC markers (Nestin) and could differentiate into neurons(β Ⅲ- tubulin) and astrocytes(GFAP). Conclusion Human fibroblasts can be converted to NSC-likes cells by 10 transcription factors.

Key words: Key words: induced neural stem/progenitor cells, human fibroblast, reprogramming, transcription factors, pax6 reporter gene

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