中国组织工程研究

中国组织工程研究 ›› 2011, Vol. 15 ›› Issue (40): 7533-7536.doi: 10.3969/j.issn.1673-8225.2011.40.028

• 干细胞基础实验 basic experiments of stem cells • 上一篇    下一篇

昆明小鼠3.5 d和4 d囊胚不同分离方法的比较

邵晓云1，徐绍业2   

  1. 桂林医学院，1组织学与胚胎学教研室，2科学实验中心，广西壮族自治区桂林市541004
  • 收稿日期:2011-04-08 修回日期:2011-05-03 出版日期:2011-10-01 发布日期:2011-10-01
  • 通讯作者: 徐绍业，讲师，桂林医学院科学实验中心，广西壮族自治区桂林市541004 shiyanshi222@163.com
  • 作者简介:邵晓云★，女，1978年生，江苏省南通市人，汉族，2004年广西大学毕业，硕士，讲师，主要从事胚胎干细胞方面的研究。 sxy2155@163.com
  • 基金资助:

    基础医学院科研启动基金(JCKY200901)，课题名称：以昆明小鼠胚胎干细胞为基础的体外心肌分化模型的初步研究。

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*

PDF

274

被引次数

4

摘要：

背景：远交系的昆明小鼠作为中国自然科学研究主要实验动物，其胚胎干细胞建系的成功率一直很低。
目的：探讨昆明小鼠胚胎干细胞体外分离培养的最佳方法和最适合的采胚时间。
方法：采集孕3.5 d和4 d的囊胚分别以免疫外科法和全胚培养法在小鼠胚胎成纤维细胞饲养层上分离和克隆昆明小鼠胚胎干细胞集落。
结果与结论：免疫外科法和全胚法培养3.5 d囊胚的内细胞团贴壁率和原代克隆形成率差异均不显著(P  > 0.05)；全胚法培养4 d囊胚的内细胞团贴壁率显著高于免疫外科法(P < 0.05)，但原代克隆形成率显著低于免疫外科法(P  < 0.05)；全胚法培养4 d囊胚的原代克隆形成率显著高于3.5 d囊胚(P  < 0.05)；免疫外科法分离4 d囊胚内细胞团的贴壁率和原代克隆形成率显著高于以同样方法分离培养的3.5 d囊胚(P  < 0.05)。结果显示用免疫外科法分离4 d囊胚更适合于昆明小鼠胚胎干细胞的体外分离培养。

关键词: 昆明小鼠, 胚胎干细胞, 全胚法, 免疫外科, 内细胞团

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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