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Modeling post-gastrula development via bidirectional pluripotent stem cells

Kuisheng Liu^1,2,† , Zihui Yan^1,2,† , Dandan Bai^1,2,† , Rui Jiang^1,2,† , Yan Bi^1,2,† , Xiangjun Ma^1,2,† , Jiani Xiang^1,2 , Yifan Sheng^1,2 , Baoxing Dong³ , Zhiyuan Ning^1,2 , Shanru Yi^1,2 , Yingdong Liu^1,2 , Xinyi Lei^1,2 , Yanping Jia^1,2 , Yan Zhang^1,2 , Yalin Zhang^1,2 , Yanhe Li^1,2 , Tao Wu^1,2 , Chenxiang Xi^1,2 , Shanyao Liu^1,2 , Shuyi Liu^1,2 , Jiayu Chen^1,2 , Jiqing Yin^1,2 , Xiaochen Kou^1,2 , Yanhong Zhao^1,2 , Hong Wang^1,2 , Yixuan Wang^1,2,* , Ke Wei^1,2,* , Shaorong Gao^1,2,* , Wenqiang Liu^1,2,*

¹Shanghai Key Laboratory of Maternal Fetal Medicine, Clinical and Translational Research Center of Shanghai First Maternity and Infant Hospital, Shanghai Institute of MaternalFetal 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
²Translational Medical Center for Stem Cell Therapy and Institute for Regenerative Medicine, Shanghai East Hospital, School of Life Sciences and Technology, Tongji University, Shanghai, China
³College of Life Sciences Technology, Shandong Normal University, Jinan, Shandong, China
^†These authors contributed equally: Kuisheng Liu, Zihui Yan, Dandan Bai, Rui Jiang, Yan Bi, Xiangjun Ma
* Correspondence: Yixuan Wang(wangyixuan@tongji.edu.cn)Ke Wei(kewei@tongji.edu.cn)Shaorong Gao(gaoshaorong@tongji.edu.cn)Wenqiang Liu(liuwenqiang@tongji.edu.cn)

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.

https://doi.org/10.1038/s41422-025-01172-x

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