Reconstructing aspects of human embryogenesis with pluripotent stem cells
Abstract
Understanding human development is of fundamental biological and clinical importance. Despite its significance, mechanisms behind human embryogenesis remain largely unknown. Here, we attempt to model human early embryo development with expanded pluripotent stem cells (EPSCs) in 3-dimensions. We define a protocol that allows us to generate self-organizing cystic structures from human EPSCs that display some hallmarks of human early embryogenesis. These structures mimic polarization and cavitation characteristic of pre-implantation development leading to blastocyst morphology formation and the transition to post-implantation-like organization upon extended culture. Single-cell RNA sequencing of these structures reveals subsets of cells bearing some resemblance to epiblast, hypoblast and trophectoderm lineages. Nevertheless, significant divergences from natural blastocysts persist in some key markers, and signalling pathways point towards ways in which morphology and transcriptional-level cell identities may diverge in stem cell models of the embryo. Thus, this stem cell platform provides insights into the design of stem cell models of embryogenesis.
Additional Information
© The Author(s) 2021. This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/. Received 27 May 2021; Accepted 24 August 2021; Published 21 September 2021. We are grateful to David Glover and Andy Cox for their helpful discussion and comments on the manuscript. We thank Min Bao for his help in single-cell sample collection; Jeff Park for his help in single-cell 10X genomics protocol and library sequencing; Matteo Guareschi for his assistance and helpful discussion in coding analysis and troubleshooting. We are grateful to Rachel S. Mandelbaum, Richard J. Paulson, Ali Ahmady and USC Fertility for their help and support to obtain images shown in Fig. 1E; Angel Martin, Maria J. de los Santos and IVIRMA Valencia for images shown in Fig. 2E. Human TSCs (TSCT) were kindly provided by Hiroaki Okae and Takahiro Arima (Tohoku University Graduate School of Medicine, Japan). The work on human embryos and stem cell models in Zernicka-Goetz Lab that supported this work was obtained from the Welcome Trust (098287/Z/12/Z), Open Philanthropy/Silicon Valley Community Foundation, Weston Havens Foundation and Shurl and Kay Curci Foundation to M.Z.G. B.S. is now funded by Yale School of Medicine. Data availability: The scRNA-seq data for 2D hEPSCs, hEP-structures, and natural human blastocyst generated in this study have been deposited in the GEO database under accession code GSE178326 [GEO]. Published iBlastoid and StemBlastoid datasets used in this study were obtained from Liu et al.20 and Yu et al.21 under accession numbers GSE156596 and GSE150578, respectively. Source data are provided with this paper. Code availability: The code generated in this study is provided at https://github.com/vjorgensen/hEP-structures_MZG. These authors contributed equally: Berna Sozen, Victoria Jorgensen. Author Contributions: B.S. designed and performed experiments and analysed the data together with V.J. M.Z. provided data on human embryos. V.J., S.C., and B.W. performed scRNA-seq data computational analysis, and statistics. B.S., V.J. B.W. and M.ZG wrote the manuscript. B.S and M.Z.G. conceived and supervised the project. Peer review information: Nature Communications thanks the anonymous reviewer(s) for their contribution to the peer review of this work.Attached Files
Published - s41467-021-25853-4.pdf
Submitted - 2021.03.12.435175v1.full.pdf
Supplemental Material - 41467_2021_25853_MOESM1_ESM.pdf
Supplemental Material - 41467_2021_25853_MOESM2_ESM.pdf
Supplemental Material - 41467_2021_25853_MOESM3_ESM.pdf
Supplemental Material - 41467_2021_25853_MOESM4_ESM.xlsx
Supplemental Material - 41467_2021_25853_MOESM5_ESM.xlsx
Supplemental Material - 41467_2021_25853_MOESM6_ESM.xlsx
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Additional details
- Alternative title
- Reconstructing human early embryogenesis in vitro with pluripotent stem cells
- PMCID
- PMC8455697
- Eprint ID
- 108455
- Resolver ID
- CaltechAUTHORS:20210317-080317393
- 098287/Z/12/Z
- Wellcome Trust
- Open Philanthropy
- Silicon Valley Community Foundation
- Weston Havens Foundation
- Shurl and Kay Curci Foundation
- Yale University
- Created
-
2021-03-19Created from EPrint's datestamp field
- Updated
-
2021-10-08Created from EPrint's last_modified field
- Caltech groups
- Division of Biology and Biological Engineering