Rolling back human pluripotent stem cells to an eight-cell embryo-like stage.
Md Abdul MazidCarl WardZhiwei LuoChuanyu LiuYunpan LiYiwei LaiLiang WuJinxiu LiWenqi JiaYu JiangHao LiuLixin FuYueli YangDavid P IbañezJunjian LaiXiaoyu WeiJuan AnPengcheng GuoYue YuanQiuting DengYang WangYing LiuFei GaoJunwen WangShahriar ZamanBaoming QinGuangming WuPatrick H MaxwellXue LiuLongqi LiuWenjuan LiMiguel Angel EstebanPublished in: Nature (2022)
After fertilization, the quiescent zygote experiences a burst of genome activation that initiates a short-lived totipotent state. Understanding the process of totipotency in human cells would have broad applications. However, in contrast to in mice 1,2 , demonstration of the time of zygotic genome activation or the eight-cell (8C) stage in in vitro cultured human cells has not yet been reported, and the study of embryos is limited by ethical and practical considerations. Here we describe a transgene-free, rapid and controllable method for producing 8C-like cells (8CLCs) from human pluripotent stem cells. Single-cell analysis identified key molecular events and gene networks associated with this conversion. Loss-of-function experiments identified fundamental roles for DPPA3, a master regulator of DNA methylation in oocytes 3 , and TPRX1, a eutherian totipotent cell homeobox (ETCHbox) family transcription factor that is absent in mice 4 . DPPA3 induces DNA demethylation throughout the 8CLC conversion process, whereas TPRX1 is a key executor of 8CLC gene networks. We further demonstrate that 8CLCs can produce embryonic and extraembryonic lineages in vitro or in vivo in the form of blastoids 5 and complex teratomas. Our approach provides a resource to uncover the molecular process of early human embryogenesis.
Keyphrases
- pluripotent stem cells
- single cell
- endothelial cells
- genome wide
- transcription factor
- dna methylation
- rna seq
- induced pluripotent stem cells
- copy number
- magnetic resonance
- high throughput
- genome wide identification
- metabolic syndrome
- pregnant women
- bone marrow
- mesenchymal stem cells
- cell free
- adipose tissue
- high frequency