Rolling back human pluripotent stem cells to an eight-cell embryo-like stage

• Published in: Nature (London) Vol. 605; no. 7909; pp. 315 - 324
• Main Authors: Mazid, Md. Abdul, Ward, Carl, Luo, Zhiwei, Liu, Chuanyu, Li, Yunpan, Lai, Yiwei, Wu, Liang, Li, Jinxiu, Jia, Wenqi, Jiang, Yu, Liu, Hao, Fu, Lixin, Yang, Yueli, Ibañez, David P., Lai, Junjian, Wei, Xiaoyu, An, Juan, Guo, Pengcheng, Yuan, Yue, Deng, Qiuting, Wang, Yang, Liu, Ying, Gao, Fei, Wang, Junwen, Zaman, Shahriar, Qin, Baoming, Wu, Guangming, Maxwell, Patrick H., Xu, Xun, Liu, Longqi, Li, Wenjuan, Esteban, Miguel A.
• Format: Journal Article
• Language: English
• Published: London Nature Publishing Group UK 12-05-2022; Nature Publishing Group
• Subjects: 13; 13/1; 13/100; 13/106; 13/31; 13/89; 14/19; 38/23; 38/39; 45; 45/41; 631/136/532/2062; 631/532/2117; Cell activation; Chromosomal Proteins, Non-Histone - genetics; Conversion; Demethylation; Deoxyribonucleic acid; DNA; DNA methylation; Embryo, Mammalian - cytology; Embryogenesis; Embryonic Development; Embryonic growth stage; Embryos; Epigenetics; Fertilization; Gametocytes; Gene expression; Genomes; Homeobox; Homeodomain Proteins - genetics; Humanities and Social Sciences; Humans; multidisciplinary; Oocytes; Pluripotency; Pluripotent Stem Cells - cytology; Production methods; Science; Science (multidisciplinary); Stem cell transplantation; Stem cells; Transcription Factors - genetics; Zygote - cytology; Zygotes
• ISSN: 0028-0836; 1476-4687
• DOI: 10.1038/s41586-022-04625-0

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
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,
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, 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
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, and TPRX1, a eutherian totipotent cell homeobox (ETCHbox) family transcription factor that is absent in mice
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. 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
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and complex teratomas. Our approach provides a resource to uncover the molecular process of early human embryogenesis.
The development of a transgene-free, rapid and controllable method for producing eight-cell-like cells from human pluripotent stem cells provides a valuable resource to study early human embryogenesis.