中国科学院广州生物医药与健康研究院 Miguel A. Esteban、Wenjuan Li、Md. Abdul Mazid和深圳华大基因Longqi Liu研究小组合作利用人类多能干细胞(PSC)产生8细胞胚胎样细胞(8CLC)。该研究于2022年3月21日发表于国际学术期刊《自然》杂志。

研究人员描述了一种无需转基因利用人类多能干细胞快速和可控产生8CLC的方法。单细胞分析确定了与这种转化相关的关键分子机制和基因网络。功能丧失实验表明DPPA3（卵母细胞中DNA甲基化的主要调节因子）和TPRX1（一种在小鼠中不存在的eutherian 全能细胞同源框(ETCHbox)家族转录因子）的基本作用。DPPA3在整个8CLC转化过程中诱导DNA去甲基化，而TPRX1是8CLC基因网络的关键基因。研究人员进一步证明8CLC可以在体外或体内以胚状体和复杂畸胎瘤的形式产生胚胎和胚胎外谱系。该方法为揭示早期人类胚胎发生的分子机制提供了一个窗口。

据悉，受精后，静止受精卵经历了基因组大量激活过程，具有短暂的全能状态。了解人类细胞的全能性具有重要的意义。然而，与小鼠相比，尚未有报道揭示体外培养人类细胞模拟合子基因组激活(ZGA)或8细胞阶段的时间图，胚胎研究受到伦理限制和实际因素的影响。

附：英文原文

Title: Rolling back of human pluripotent stem cells to an 8-cell embryo-like stage

Author: 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, Ibaez, 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.

Issue&Volume: 2022-03-21

Abstract: Upon fertilization, the quiescent zygote experiences a burst of genome activation that initiates a short-lived totipotent state. Understanding totipotency in human cells would have very broad implications. However, in contrast to mouse1,2, proper demonstration of in vitro cultured human cells mimicking the time of zygotic genome activation (ZGA) or 8-cell (8C) stage had 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 (8CLC) from human pluripotent stem cells (PSC). Single-cell analysis identified key molecular events and gene networks associated with this conversion. Loss-of-function experiments showed fundamental roles for DPPA3, a master regulator of DNA methylation in oocytes3, and TPRX1, a eutherian totipotent cell homeobox (ETCHbox) family transcription factor absent in mouse4. DPPA3 induces DNA demethylation throughout the 8CLC conversion, whereas TPRX1 is a key executor of the 8CLC gene networks. We further demonstrate that 8CLC can produce embryonic and extraembryonic lineages in vitro or in vivo in the form of blastoids5 and complex teratomas. Our approach provides a window to uncover the molecular logic of early human embryogenesis.

DOI: 10.1038/s41586-022-04625-0

Source: https://www.nature.com/articles/s41586-022-04625-0