四倍体滋养层干细胞的分离培养与鉴定

Isolation and identification of tetraploid trophoblast stem cells

目的建立可在体外培养的小鼠四倍体滋养层干细胞(TTSCs)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,挑取克隆并传代后获得细胞系。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TTSCs。分化条件下,TTSCs分化基因的表达量与二倍体滋养层干细胞(TSCs)存在差异(P<0.001)。结论可传代的TTSCs系可以在体外建立,但其维持自我更新及分化的机制与二倍体TSCs有所不同。

Objective To establish mice tetraploid trophoblast stem cell(TTSCs)line 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 TTSCs that can develop in vivo and be passaged were established.The expression of differentiation genes was distinct between TTSCs diploid TSCs under differentiation condition.Conclusions Passable cell line TTSCs could be established in vitro,but the mechanisms of self-renew and differentiation were different between TTSCs and diploid TSCs.