解剖学报

›› 2012, Vol. 43 ›› Issue (5): 601-607.doi: 10.3969/j.issn.0529-1356.2012.05.004

• 细胞和分子生物学 • 上一篇    下一篇

人包皮成纤维细胞来源的多能性干细胞的建立

李雪1; 单智焱1; 吴嫣爽1; 申景岭1; 杨玉芝2; 雷蕾1*   

  1. 1.哈尔滨医科大学组织学与胚胎学教研室，哈尔滨 150081; 2.黑龙江省医院内分泌科，哈尔滨 150036
  • 收稿日期:2012-01-30 修回日期:2012-03-29 出版日期:2012-10-06
  • 通讯作者: 雷蕾

Generation of induced pluripotent stem cells from human foreskin fibroblasts

  1. 1.Department of Histology and Embryology, Harbin Medical University, Harbin 150081, China; 2. Department of Endocrinology,Heilongjiang Province Hospital, Harbin 150036, China
  • Received:2012-01-30 Revised:2012-03-29 Online:2012-10-06
  • Contact: LEI Lei

关键词: 干细胞, 分化, 包皮成纤维细胞, 绿色荧光蛋白, 免疫荧光, 人

Abstract: Objective To optimize the method for establishment of induced pluripotent stem cells from human foreskin fibroblasts. Methods We used GFP as an indicator to determine the optimized dose of the retrovirus during the reprogramming of human foreskin fibroblasts into human induced pluripotent stem cells (iPSCs) by Yamanaka’s classical strategy with the combination of the small-molecule compounds VPA, a histone deacetylase inhibitor. The characteristics of established iPSCs were tested via alkaline phophatase (AP) staining, immunofluorescent detection of human embryonic stem (ES) cells surface antigens, and RT-PCR analysis of three germ layers specific gene expression in cell during differentiation in vitro. Results Using GFP expression as an indicator of the retrovirus transduction, we established a high efficient system to successfully reprogramme human foreskin fibroblasts into induced pluripotent stem cells. The iPSCs showed positive for alkaline phosphatase, expressed high levels of human ES cells pluripotency markers such as OCT4,TRA-1-60 and TRA-1-81. RT-PCR analysis indicated that the germ layers differentiation related gene expression was detected in the embryonic body when derived from these hiPS cells. BR>Conclusion Our results showed that we successfully generated hiPS cells from adult foreskin fibroblasts in the indication of GFP with the combination of the small-molecule compoun

Key words: Stem cell, Differentiation, Foreskin fibroblast, Green fluorescent protein, Immunofluorescence, Human

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