解剖学报

›› 2009, Vol. 40 ›› Issue (3): 409-413.doi: 10.3969/j.issn.0529-1356.2009.03.013

• 论著 • 上一篇    下一篇

不同类型的转基因细胞核供体对生产小鼠转基因克隆胚胎的影响

闫晓飞1 ;徐燕宁1; 李洪武2 ;关娜1; 单智焱1 ;钟淑琦1; 金连弘1 ;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.哈尔滨医科大学第一临床医学院立体定向功能神经外科,哈尔滨 150001
  • 收稿日期:2008-01-07 修回日期:2008-04-14 出版日期:2009-06-06
  • 通讯作者: 雷蕾

Production of transgenic cloned mouse embryos by somatic cell nuclear transfer with different donor cells

  1. 1.Department of Histology and Embryology， Harbin Medical University， Harbin 150081, China;2.Deparment of Stereotactic and Functional Neurosurgery,the First Affiliated Hospital of Harbin Medical University,Harbin 150001,China
  • Received:2008-01-07 Revised:2008-04-14 Online:2009-06-06
  • Contact: LEI Lei

关键词: 核移植, 转基因体细胞, 电穿孔, 绿色荧光蛋白, 小鼠

Abstract: Objective To generate transgenic mouse embryos by somatic cell nuclear transfer (SCNT), and compare effects of different donor cells on developmental potential of cloned embryos. Methods Mouse embryo fibroblasts and mouse embryonic stem (ES) cells were transfected with a plasmid (pCE-EGFP-IRES-NEO) containing the enhanced green fluorescent protein (GFP) and neomycinresistant (Neor) genes by electroporation. Transgenic cell lines were obtained for SCNT after 14 days selection with G418. At the same time, SCNT was performed using cumulus cells and normal fibroblasts as control. Results There was no significant difference in blastocyst rates between reconstructed embryos derived from transgenic fibroblasts and nontransgenic ones (154/160,19.23% vs 22.91%, EM>P/EM>＞0.05). Reconstructed embryos from transgenic ES cells showed higher blastocyst developmental rates than that from transgenic fibroblasts donor (152/154,41.54% vs 19.23%, P<0.05). Cumulus cloned embryos had better developmental potential than that of fibroblasts cloned ones (171/160, 41.17% vs 22.91%, P<0.05). Conclusion Our results showed that transgenic donor cells did not affect EM>in vitro/EM> developmental potential of mouse SCNT embryos. Using EGFP maker, mouse transgenic blastocysts could be produced effectively

Key words: Nuclear transfer, Transgenic somatic cell, Electroporation, green flourescent protein, Mouse

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