Liu Kuisheng, Yan Zihui, Bai Dandan, Jiang Rui, Bi Yan, Ma Xiangjun, Xiang Jiani, Sheng Yifan, Dong Baoxing, Ning Zhiyuan, Yi Shanru, Liu Yingdong, Lei Xinyi, Jia Yanping, Zhang Yan, Zhang Yalin, Li Yanhe, Wu Tao, Xi Chenxiang, Liu Shanyao, Liu Shuyi, Chen Jiayu, Yin Jiqing, Kou Xiaochen, Zhao Yanhong, Wang Hong, Wang Yixuan, Wei Ke, Gao Shaorong, Liu Wenqiang
Shanghai Key Laboratory of Maternal Fetal Medicine, Clinical and Translational Research Center of Shanghai First Maternity and Infant Hospital, Shanghai Institute of Maternal-Fetal Medicine and Gynecologic Oncology, Frontier Science Center for Stem Cell Research, Shanghai Key Laboratory of Signaling and Disease Research, School of Life Sciences and Technology, Tongji University, Shanghai, China.
Translational Medical Center for Stem Cell Therapy and Institute for Regenerative Medicine, Shanghai East Hospital, School of Life Sciences and Technology, Tongji University, Shanghai, China.
Cell Res. 2025 Aug 29. doi: 10.1038/s41422-025-01172-x.
The absence of stem cells capable of efficiently generating both trophoblast and epiblast lineages has hindered precise recapitulation of embryonic development. Through high-content chemical screening, we established an (AS and LY) AL medium to generate mouse bidirectional pluripotent stem cells (BPSCs) characterized by concurrent expression of OCT4 and CDX2. Mouse BPSCs demonstrated highly plastic differentiation into trophoblast, epiblast and primitive endoderm (PrE) lineages in vitro within 48 h without exogenous inducing factors and efficiently contributed to embryonic and extraembryonic tissues in vivo. Mechanistically, hyperactivation of the Wnt signaling pathway breaks the early lineage differentiation barrier by initiating a Lef1-dependent bypass. Remarkably, integration of BPSCs with PrE induction system enables high-efficiency generation of E8.5-stage embryo models. These advanced models complete gastrulation and recapitulate definitive developmental milestones including brain morphogenesis, neural tube closure, cardiac contraction, somite patterning, and primordial germ cell specification. Moreover, human cells cultured under AL conditions acquire an OCT4 and CDX2 double-positive state and corresponding gene expression profiles, revealing conserved functionality of this culturing platform across species. These findings highlight BPSCs as a powerful tool for investigating early lineage specification and post-gastrulation embryonic development.
缺乏能够有效产生滋养层和上胚层谱系的干细胞阻碍了对胚胎发育的精确重现。通过高内涵化学筛选，我们建立了一种（AS和LY）AL培养基，以生成具有OCT4和CDX2共表达特征的小鼠双向多能干细胞（BPSC）。小鼠BPSC在无外源性诱导因子的情况下，于48小时内在体外高度可塑性地分化为滋养层、上胚层和原始内胚层（PrE）谱系，并在体内有效地参与胚胎和胚外组织的形成。从机制上讲，Wnt信号通路的过度激活通过启动Lef1依赖性旁路打破了早期谱系分化障碍。值得注意的是，将BPSC与PrE诱导系统整合可高效生成E8.5期胚胎模型。这些先进模型完成了原肠胚形成，并重现了包括脑形态发生、神经管闭合、心脏收缩、体节模式形成和原始生殖细胞特化在内的确定发育里程碑。此外，在AL条件下培养的人类细胞获得了OCT4和CDX2双阳性状态及相应的基因表达谱，揭示了该培养平台在物种间的保守功能。这些发现突出了BPSC作为研究早期谱系特化和原肠胚形成后胚胎发育的有力工具。
