解剖学报

›› 2011, Vol. 42 ›› Issue (2): 279-282.doi: 10.3969/j.issn.0529-1356.2011.02.028

• 论著 • 上一篇    下一篇

无血清无饲养层人胚胎干细胞向胶质前体细胞的分化

罗敏1,3;胡智兴2,3*;梁道明4   

  1. 1.昆明医学院药学院暨云南省天然药物药理重点实验室;2.药理学教研室;3. 云南省生物医学工程中心，昆明650500;4. 昆明医学院第二附属医院，昆明650031
  • 收稿日期:2010-04-12 修回日期:2010-10-07 出版日期:2011-04-06
  • 通讯作者: 胡智兴

Derivation of neuroglia progenitors from human embryonic stem cells in feeder layer- and serum-free medium

  1. 1. Faculty of Pharmacy & Yunnan Key Laboratory of Pharmacology for Natural Product; 2. Department of Pharmacology; 3. Biomedical Engineering Research Centre of Yunnan University,Kunming Medical University, Kunming650500, China; 4. The Second Affiliated Hospital, Kunming Medical University, Kunming650031, China
  • Received:2010-04-12 Revised:2010-10-07 Online:2011-04-06
  • Contact: HU Zhi-xing

关键词: 胚胎干细胞, 胶质前体细胞, 免疫荧光, 反转录-聚合酶链反应, 人

Abstract: Objective To establish a neuroglia progenitor differentiation system from human embryonic stem cells (hESCs) in a feeder layer- and serum-free medium. Methods hESCs grown in laminin coated culture plates were induced to form embryoid bodies (EBs) for 25 days. Then 25-day-old EBs were transferred to glial progenitor induction medium (GPIM) containing insulin, 5μg/L basic fibroblast growth factor (bFGF), 20μg/L epidermal growth factor (EGF) and 5μg/L triiodothyronine (T3) to continue culturing for 7-10 days. The differentiated fibroblast-like cells were digested and collected. The expressions of neuroglia progenitor genes were determined by RT-PCR. The special markers of neuroglia progenitors were examined by immunofluorescence staining. Results The differentiated cells were long spindle shaped. They expressed neuroglia progenitor special markers NG2, platelet\|derived growth factor receptor α（PDGFRα）. The percentage of NG2+, PDGFRα+ cells was 37.2% and 47.6%, respectively. They could further differentiate into astrocytes and oligodendroglias.Conclusion GPIM containing insulin, bFGF, EGF and T3 could induce hESCs differentiate into neuroglia progeni

Key words: Embryonic stem cell, Glial progenitor, Immunofluorescence, RT-PCR, Human

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