基础医学与临床 ›› 2021, Vol. 41 ›› Issue (7): 975-981.

• 研究论文 • 上一篇    下一篇

四倍体滋养层干细胞的分离培养与鉴定

王海豫,肖翼,黄粤   

  1. 中国医学科学院基础医学研究所
  • 收稿日期:2021-04-06 修回日期:2021-05-18 出版日期:2021-07-05 发布日期:2021-06-17
  • 通讯作者: 黄粤 E-mail:huangyue@pumc.edu.cn
  • 基金资助:
    全能性细胞调节因子的遗传筛选和培养体系的建立。

Isolation and identification of tetraploid trophoblast stem cells

  • Received:2021-04-06 Revised:2021-05-18 Online:2021-07-05 Published:2021-06-17
  • Contact: Huang Yue E-mail:huangyue@pumc.edu.cn

摘要: 目的 建立可在体外培养的小鼠四倍体滋养层干细胞系,为研究胎盘发育提供新的细胞模型。方法 用电融合的方法获得四倍体胚胎并进行体外培养,挑取克隆后传代后获得细胞系。通过中期染色体计数检测染色体数目,RT-qPCR、Western blot、免疫荧光检测标志基因表达,显微注射检测细胞的体内发育潜能。结果 建立了可以传代培养并且可在体内发育的四倍体滋养层干细胞系(tetraploid trophoblast stem cells, TTSCs)。分化条件下,TTSCs分化基因的表达量与二倍体TSCs存在差异。结论 TTSCs可以在体外建立,但其维持自我更新及分化的机制与二倍体TSCs有所不同。

关键词: 四倍体, 滋养层干细胞, 胚胎发育, 胎盘发育, 分化

Abstract: Objective To establish mice tetraploid trophoblast stem cell lines which could be cultured in vitro, providing a new cell model for the research of placenta development. Methods Tetraploid embryos were produced by electrofusion and cultured in vitro, clones were picked up and passaged to establish cell lines. Metaphase chromosome counting was used to detect the chromosome number. RT-qPCR, western blot and immunofluorescence were used to detect the expression of marker genes. Microinjection was used to detect the development potential in vivo. Results Tetraploid trophoblast stem cell lines (TTSCs) that can develop in vivo and be passaged could be established. The expression of differentiation genes was distinct between TTSCs and diploid TSCs under differentiation condition. Conclusions TTSCs could be established in vitro, but the mechanisms of self-renew and differentiation were different between TTSCs and diploid TSCs.

Key words: tetraploid, trophoblast stem cell, embryogenesis, placenta development, differentiation

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