Chinese Journal of Tissue Engineering Research ›› 2011, Vol. 15 ›› Issue (40): 7533-7536.doi: 10.3969/j.issn.1673-8225.2011.40.028

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Comparison of different methods to isolate 3.5-day and 4-day blastocysts from Kunming mice

Shao Xiao-yun1, Xu Shao-ye2   

  1. 1Department of Histology and Embryology, 2Science and Technology Division, Guilin Medical University, Guilin  541004, Guangxi Zhuang Autonomous Region, China
  • Received:2011-04-08 Revised:2011-05-03 Online:2011-10-01 Published:2011-10-01
  • Contact: Xu Shao-ye, Lecturer, Science and Technology Division, Guilin Medical University, Guilin 541004, Guangxi Zhuang Autonomous Region, China shiyanshi222@163.com
  • About author:Shao Xiao-yun★, Master, Lecturer, Department of Histology and Embryology, Guilin Medical University, Guilin 541004, Guangxi Zhuang Autonomous Region, China sxy2155@163.com
  • Supported by:

    the Science and Technology Research Startup Fund of Basic Medical School, No. JCKY200901*

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Abstract:

BACKGROUND: Outbreed Kunming mice as the main experimental animals for natural science research in China have a low successful rate in establishment of embryonic stem cell line.
OBJECTIVE: To explore the method of isolation and culture of murine embryonic stem cells and the best time to collect embryos of Kunming species.
METHODS: Blastocysts obtained from 3.5-day and 4-day embryos after mating were treated with intact embryo cultivation and immuosurgery. Inner cell masses were isolated and cultured on a feeder layer of mouse embryo fobroblasts (MEF). And colony growth was observed.
RESULTS AND CONCLUSION: Adherent rate of inner cell mass by inatact embryo cultural method were higher than those by immunosurgical method from 4-day (P< 0.05), but in the rate of primary colony formation, the latter was better than the former. And, in the rate of primary colony formation, 4-day was better than 3.5-day by intact embryo culture method (P < 0.05). By immunosurgical method, adherent rate of inner cell mass and the rate of primary colony formation from 4-day were higer than those of 3.5-day (P < 0.05). Immunosurgical method to isolate 4-day embryos is helpful to culture embryonic stem cells of Kunming mice in vitro.

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