Transgene-free generation of mouse post-gastrulation whole embryo models solely from naive ESCs and iPSCs

  • Cell Stem Cell. 2025 Aug 7:S1934-5909(25)00262-0. doi: 10.1016/j.stem.2025.07.005.
Alperen Yilmaz  1 ,  Gulben Gurhan  2 ,  Mehmet-Yunus Comar  3 ,  Sergey Viukov  3 ,  Inbal Serfaty  3 ,  Mert Gayretli  3 ,  Sergey Golenchenko  3 ,  Dmitry Lokshtanov  3 ,  Shahd Ashouokhi  3 ,  Angel Polanco  3 ,  Idan Berlad  3 ,  Tae-Won Ha  3 ,  Alejandro Aguilera-Castrejon  4 ,  Shadi Tarazi  3 ,  Marina Cohen  5 ,  Nir Livnat  3 ,  Komal Kumar  6 ,  Hisham Cholakkal  6 ,  Nathan Levy  7 ,  Nir Yosef  7 ,  Nizar Khatib  8 ,  Reli Rachel Kakun  9 ,  Merav Kedmi  10 ,  Inbal Bolocan Nachman  10 ,  Hadas Keren-Shaul  10 ,  Yoseph Addadi  10 ,  Ayelet-Hashahar Orenbuch  11 ,  Karina Korovin  12 ,  Alina Molchadsky  12 ,  Konrad Hochedlinger  13 ,  Ohad Gafni  12 ,  Itay Maza  14 ,  Noa Novershtern  15 ,  Bernardo Oldak  16 ,  Jacob H Hanna  17
Affiliations
  • 1. Department of Molecular Genetics, Weizmann Institute of Science, Rehovot, Israel. Electronic address: [email protected].
  • 2. Department of Molecular Genetics, Weizmann Institute of Science, Rehovot, Israel. Electronic address: [email protected].
  • 3. Department of Molecular Genetics, Weizmann Institute of Science, Rehovot, Israel.
  • 4. Janelia Research Campus, Howard Hughes Medical Institute, Ashburn, VA, USA.
  • 5. Department of Veterinary Resources, Weizmann Institute of Science, Rehovot, Israel.
  • 6. Mohamed bin Zayed University of Artificial Intelligence (MBZUAI), Abu-Dhabi, UAE.
  • 7. Department of Systems Immunology, Weizmann Institute of Science, Rehovot, Israel.
  • 8. Department of Obstetrics and Gynecology, Rambam Health Care Campus, Haifa, Israel; Rappaport Faculty of Medicine, Technion Israel Institute of Technology, Haifa, Israel.
  • 9. Clinical Research Institute at Rambam, Rambam Health Care Campus, Haifa, Israel.
  • 10. Department of Life Sciences Core Facilities, Weizmann Institute of Science, Rehovot, Israel.
  • 11. Department of Molecular and Cell Biology, Weizmann Institute of Science, Rehovot, Israel.
  • 12. RenewalBio LTD, Rehovot, Israel.
  • 13. Department of Molecular Biology, Massachusetts General Hospital, Harvard Medical School, Boston, MA, USA.
  • 14. Rappaport Faculty of Medicine, Technion Israel Institute of Technology, Haifa, Israel; Gastroenterology Unit, Rambam Health Care Campus, Haifa, Israel.
  • 15. Department of Molecular Genetics, Weizmann Institute of Science, Rehovot, Israel. Electronic address: [email protected].
  • 16. Department of Molecular Genetics, Weizmann Institute of Science, Rehovot, Israel. Electronic address: [email protected].
  • 17. Department of Molecular Genetics, Weizmann Institute of Science, Rehovot, Israel. Electronic address: [email protected].
PMID: 40780191 DOI: 10.1016/j.stem.2025.07.005
Abstract

The generation of post-gastrulation stem cell-derived mouse embryo models (SEMs) exclusively from naive embryonic stem cells (nESCs) has underscored their ability to give rise to embryonic and extra-embryonic lineages. However, existing protocols for mouse SEMs rely on the separate induction of extra-embryonic lineages and on ectopic expression of transcription factors to induce nESC differentiation into trophectoderm (TE) or primitive endoderm (PrE). Here, we demonstrate that mouse nESCs and naive induced pluripotent stem cells (niPSCs) can be simultaneously co-induced, via signaling pathway modulation, to generate PrE and TE extra-embryonic cells that self-organize into embryonic day (E) 8.5-E8.75 transgene-free (TF) SEMs. We also devised an alternative condition (AC) naive media that in vitro stabilizes TF-SEM-competent OCT4+/NANOG+ nESC colonies that co-express antagonistic CDX2 and/or GATA6 extra-embryonic fate master regulators and self-renew while remaining poised for TE and PrE differentiation, respectively. These findings improve mouse SEM strategies and shed light on amplifying an inherent and dormant extra-embryonic plasticity of mouse naive pluripotent cells in vitro.

Keywords
chemical reprogramming; embryo models; ex utero embryo culture; extra-embryonic lineages; naive pluripotency; primitive endoderm; synthetic embryology; trophoblast stem cells.
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