人诱导性多能干细胞定向分化为神经干细胞的实验研究

基础医学与临床 ›› 2010, Vol. 30 ›› Issue (12): 1263-1267.

人诱导性多能干细胞定向分化为神经干细胞的实验研究

冯年花¹,谢安¹,娄远蕾²,阮琼芳²,郭菲²,汪泱²

1. 南昌大学第一附属医院泌尿外科研究所2. 南昌大学一附院泌尿外科研究所

收稿日期:2009-11-17 修回日期:2010-03-30 出版日期:2010-12-05 发布日期:2010-12-05
通讯作者: 汪泱

Differentiation of human induced pluripotent stem cells into neural stem cells

FENG Nian-hua,XIE An ,LOU Yuan-lei ,RUAN Qiong-fang ,GUO Fei ,WANG Yang

Nanchang University

Received:2009-11-17 Revised:2010-03-30 Online:2010-12-05 Published:2010-12-05
Contact: WANG Yang

摘要/Abstract

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-TUBULIN Ш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-TUBULIN Ш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. The transformation of cell morphology 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-TUBULINШ, 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-TUBULINШ positive neurons and few GFAP positive astrocytes. There were no neural tube-like constructions in control group. Conclusion Human iPS cells can differentiate toward neural stem cells and can imitate the development process of nervous system.

冯年花;谢安;娄远蕾;阮琼芳;郭菲;汪泱. 人诱导性多能干细胞定向分化为神经干细胞的实验研究[J]. 基础医学与临床, 2010, 30(12): 1263-1267.

FENG Nian-hua;XIE An ;LOU Yuan-lei ;RUAN Qiong-fang ;GUO Fei ;WANG Yang. Differentiation of human induced pluripotent stem cells into neural stem cells[J]. Basic & Clinical Medicine, 2010, 30(12): 1263-1267.