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Published January 14, 2026 | Version Published
Journal Article Open

Efficient stem cell-derived mouse embryo models for environmental studies

Creators

  • 1. ROR icon California Institute of Technology
  • 2. ROR icon University of Basel
  • 3. ROR icon Kaiser Permanente
  • 4. ROR icon University of Cambridge

Abstract

Blastoids are stem cell-derived structures that mimic natural blastocysts by incorporating all three lineages: trophectoderm, epiblast, and primitive endoderm. However, current methods often yield incomplete structures that fail to cavitate or to form a proper primitive endoderm. To overcome these limitations, we develop a modular approach by aggregating three murine stem cell types: embryonic stem cells (ESCs), ESCs with inducible GATA4 expression (iG4-ESCs), and trophoblast stem cells (TSCs). This method yields cavitated blastocyst-like structures-termed iG4-blastoids-with approximately 80% efficiency. Single-cell RNA sequencing confirms their close resemblance to mature mouse blastocysts. Notably, culturing iG4-blastoids without FGF4 enhances specification of the invasive mural trophectoderm, and approximately 12% of structures undergo post-implantation-like morphogenesis in vitro. Using this model, we show that caffeine, alcohol, nicotine, and amino acid variations affect iG4-blastoids and natural embryos similarly, underscoring their utility as a robust model for investigating the impact of diverse environmental factors on embryogenesis.

Copyright and License

© 2025 The Authors. Published by Elsevier Inc. This article is available under the Creative Commons CC-BY-NC-ND license and permits non-commercial use of the work as published, without adaptation or alteration provided the work is fully attributed.

Acknowledgement

We thank Angela Andersen and the past and present members of the Zernicka-Goetz lab for invaluable advice during the preparation of this manuscript. Sequencing was performed at the UCSF CAT, supported by UCSF PBBR, RRP IMIA, and NIH 1S10OD028511-01 grants. This work was supported by the National Institute of Health via the Director’s Pioneer Award (5DP1HD104575-04 to M.Z.-G.), the Open Philanthropy Award (M.Z.-G.), and R01HD101489 to M.Z.-G., the Human Frontiers Science Program (LT-0022/2022 to S.J.), the NSFC (82301873 to M.B.), the Ministry of Science and Technology of China (2024YFA1107002 to M.B.), the Swiss Study Foundation (C.M.H.), the Werenfels Fund of the Free Academic Society (Freie Akademische Gesellschaft) Basel (C.M.H.), and the Claudine and Hans-Heiner Zaeslin-Bustany Foundation (C.M.H.).

Supplemental Material

Supplemental files attached:

  • Document S1. Figures S1–S6 and Tables S1–S6.
  • Document S2. Article plus supplemental information.

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Additional details

Identifiers

PMID
40882624

Funding

National Natural Science Foundation of China
82301873
Ministry of Science and Technology of the People's Republic of China
2024YFA1107002
International Human Frontier Science Program Organization
LT-0022/2022
National Institutes of Health
1S10OD028511-01
National Institutes of Health
Director’s Pioneer Award 5DP1HD104575-04
International Human Frontier Science Program Organization
LT-0022/2022
National Institutes of Health
R01HD101489

Dates

Available
2025-08-28
Published online
Available
2026-01-14
Version of record

Caltech Custom Metadata

Caltech groups
Division of Biology and Biological Engineering (BBE)
Publication Status
Published