A model of the post-implantation human embryo derived from pluripotent stem cells

Creators: Weatherbee, Bailey A. T.¹; Gantner, Carlos W.¹; Iwamoto-Stohl, Lisa K.¹; Daza, Riza M.²; Hamazaki, Nobuhiko²; Shendure, Jay^{2, 3, 4}; Zernicka-Goetz, Magdalena^{1, 2, 5}

1. University of Cambridge
2. University of Washington
3. Brotman Baty Institute
4. Howard Hughes Medical Institute
5. California Institute of Technology

Abstract

The human embryo undergoes morphogenetic transformations following implantation into the uterus, yet our knowledge of this crucial stage is limited by the inability to observe the embryo in vivo. Stem cell-derived models of the embryo are important tools to interrogate developmental events and tissue-tissue crosstalk during these stages. Here, we establish a model of the human post-implantation embryo, a human embryoid, comprised of embryonic and extraembryonic tissues. We combine two types of extraembryonic-like cells generated by transcription factor overexpression with wildtype embryonic stem cells and promote their self-organization into structures that mimic several aspects of the post-implantation human embryo. These self-organized aggregates contain a pluripotent epiblast-like domain surrounded by extraembryonic-like tissues. Our functional studies demonstrate that the epiblast-like domain robustly differentiates to amnion, extraembryonic mesenchyme, and primordial germ cell-like cells in response to BMP cues. In addition, we identify an inhibitory role for SOX17 in the specification of anterior hypoblast-like cells. Modulation of the subpopulations in the hypoblast-like compartment demonstrated that extraembryonic-like cells impact epiblast-like domain differentiation, highlighting functional tissue-tissue crosstalk. In conclusion, we present a modular, tractable, integrated model of the human embryo that will allow us to probe key questions of human post-implantation development, a critical window when significant numbers of pregnancies fail.

Additional Information

© The Author(s), under exclusive licence to Springer Nature Limited 2023. The authors are grateful to CARE Fertility, Herts and Essex Fertility Centre, Bourn Hall Fertility Clinic, and King's Fertility Clinic for their collaboration in the donation of supernumerary human embryos. The authors thank all members of the M.Z-G and Boroviak labs, Gianluca Amadei and David Glover for feedback, Marta Shahbazi for gifting the Shef6-mKate2 cell line and for feedback on the manuscript. This work is supported by Wellcome Trust (207415/Z/17/Z), in part by European Research Council (669198), Open Atlas, and NOMIS award grants to M.Z-G, Allen Discovery Center for Cell Lineage Tracing grants to J.S. and M.Z-G., in addition to individual funding from the Gates Cambridge Trust (to B.A.T.W.) and Leverhulme Trust Early Career Fellowship (to C.W.G.). J.S. is an investigator of the Howard Hughes Medical Institute and M.Z966 G. is NOMIS Distinguished Scientist and Scholar. Author Contributions. B.A.T.W. and C.W.G. designed, carried out, and analyzed experiments. B.A.T.W. and C.W.G. independently reproduced human embryoid generation and results. B.A.T.W., C.W.G. and L.K.I-S. performed human embryo work. L.K.I-S performed mouse embryo-hPSC chimera experiments. N.H. and R.M.D. prepared 10x multiome libraries and performed RNA and ATAC sequencing supervised by J.S. B.A.T.W. analyzed single cell sequencing data. C.W.G. and B.A.T.W. wrote the manuscript with input from all co-authors. M.Z-G conceived and supervised the project. Declarations of Interests. Authors are inventors on the following patents: 1. Patent Applicant: Caltech. Inventors: Magdalena Zernicka-Goetz, Berna Sozen, Victoria Jorgensen. Application Number: 17/692,790. Specific aspect of the manuscript covered in patent application: Reconstructing Human Early Embryogenesis In Vitro With Pluripotent Stem Cells. 2. Patent Applicant: Caltech and Cambridge Enterprise Limited. Inventors: Magdalena Zernicka-Goetz, Gianluca Amadei, Charlotte Handford. Application Number: 63/397,630 Specific aspect of the manuscript covered in patent application: Synthetic Embryos. 3. Patent Applicant: Caltech and Cambridge Enterprise Limited; Inventors: Magdalena Zernicka-Goetz, Bailey Weatherbee, Carlos Gantner. Application Number: 63/403,684 Specific aspect of the manuscript covered in patent application: Stem Cell Derived Model Of The Human Embryo. Data Availability. For aligning sequencing data, GRCh38 (https://www.ncbi.nlm.nih.gov/assembly/GCF_000001405.26/) and GRCm38 (https://www.ncbi.nlm.nih.gov/assembly/GCF_000001635.20/) were used. Previously published data is publicly available: Human data: Molè et al., 2021: ArrayExpress E-MTAB-8060 Xiang et al., 2020: Gene Expression Omnibus GSE136447 Zhou et al., 2019: Gene Expression Omnibus GSE109555 Petropoulos et al., 2016: ArrayExpress E-MTAB-3929 Blakely et al., 2015: Gene Expression Omnibus GSE66507 Yan et al., 2013: Gene Expression Omnibus GSE36552 Cynomolgus Monkey: Yang et al., 2021: Gene Expression Omnibus GSE148683 Ma et al., 2019: Gene Expression Omnibus GSE130114 Nakamura et al., 2016: Gene Expression Omnibus GSE74767 hESC derived datasets: Pham et al., 2022: Gene Expression Omnibus GSE191286 Kagawa et al., 2022: Gene Expression Omnibus GSE177689 Zheng et al., 2019: Gene Expression Omnibus GSE134571 10x multiome data for cell lines and inducible human embryoids: Gene Expression Omnibus GSE218314 Source data are provided with this paper. Code Availability. No custom code was utilized in this manuscript. Code used to analyze these data is available at https://github.com/bweatherbee/human_model.

Attached Files

Accepted Version - s41586-023-06368-y_reference.pdf

Files

s41586-023-06368-y_reference.pdf

Files (33.6 MB)

Name	Size	Download all
s41586-023-06368-y_reference.pdf md5:6e1615c44fde9067ae19354b208d0a22	33.6 MB	Preview Download

Additional details

	All versions	This version
Views	79	79
Downloads	18	18
Data volume	605.6 MB	605.6 MB