biostudies-literatureUnknown626(7998)Okubo TRecently, several studies using cultures of human embryos together with single-cell RNA-seq analyses have revealed differences between humans and mice, necessitating the study of human embryos^1-8. Despite the importance of human embryology, ethical and legal restrictions have limited post-implantation-stage studies. Thus, recent efforts have focused on developing in vitro self-organizing models using human stem cells^9-17. Here, we report genetic and non-genetic approaches to generate authentic hypoblast cells (naive hPSC-derived hypoblast-like cells (nHyCs))-known to give rise to one of the two extraembryonic tissues essential for embryonic development-from naive human pluripotent stem cells (hPSCs). Our nHyCs spontaneously assemble with naive hPSCs to form a three-diNature357-366https://www.ebi.ac.uk/biostudies/studies/S-EPMC10849967biostudies-literatureHypoblast from human pluripotent stem cells regulates epiblast development.PMC10849967Woltjen KRivron NKishimoto KYamamoto TSaitou MNakajima-Koyama MKabata MNakauchi HTakashima YMasaki HOkubo TSasaki EKaswandy BSemi KSato HKunitomi HfalseHypoblast from human pluripotent stem cells regulates epiblast development.Recently, several studies using cultures of human embryos together with single-cell RNA-seq analyses have revealed differences between humans and mice, necessitating the study of human embryos^1-8. Despite the importance of human embryology, ethical and legal restrictions have limited post-implantation-stage studies. Thus, recent efforts have focused on developing in vitro self-organizing models using human stem cells^9-17. Here, we report genetic and non-genetic approaches to generate authentic hypoblast cells (naive hPSC-derived hypoblast-like cells (nHyCs))-known to give rise to one of the two extraembryonic tissues essential for embryonic development-from naive human pluripotent stem cells (hPSCs). Our nHyCs spontaneously assemble with naive hPSCs to form a three-di2024-01-01T00:00:00Z2024 Feb2026-04-29T14:08:54.804Z2025-02-18T23:14:02.593ZS-EPMC108499673805222810.1038/s41586-023-06871-2