High-resolution transcriptional and morphogenetic profiling of cells from micropatterned human ESC gastruloid cultures
- Washington University in St Louis, United States
- Washington University School of Medicine, United States
- University of California, Davis, United States
- University of Illinois, Urbana-Champaign, United States
Abstract
During mammalian gastrulation, germ layers arise and are shaped into the body plan while extraembryonic layers sustain the embryo. Human embryonic stem cells, cultured with BMP4 on extracellular matrix micro-discs, reproducibly differentiate into gastruloids, expressing markers of germ layers and extraembryonic cells in radial arrangement. Using single-cell RNA sequencing and cross-species comparisons with mouse, cynomolgus monkey gastrulae, and post-implantation human embryos, we reveal that gastruloids contain cells transcriptionally similar to epiblast, ectoderm, mesoderm, endoderm, primordial germ cells, trophectoderm, and amnion. Upon gastruloid dissociation, single cells reseeded onto micro-discs were motile and aggregated with the same but segregated from distinct cell types. Ectodermal cells segregated from endodermal and extraembryonic but mixed with mesodermal cells. Our work demonstrates that the gastruloid system models primate-specific features of embryogenesis, and that gastruloid cells exhibit evolutionarily conserved sorting behaviors. This work generates a resource for transcriptomes of human extraembryonic and embryonic germ layers differentiated in a stereotyped arrangement.
Data availability
Sequencing data have been deposited in GEO under accession code GSE144897.
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In vitro culture of cynomolgus monkey embryos beyond gastrulationNCBI Gene Expression Omnibus, GSE130114.
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A developmental coordinate of the spectrum of pluripotency among mice, monkeys, and humansNCBI Gene Expression Omnibus, GSE74767.
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A developmental landscape of 3D-cultured human pre-gastrulation embryosNCBI Gene Expression Omnibus, GSE136447.
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Human Primordial Germ Cells are Specified from Lineage Primed ProgenitorsNCBI Gene Expression Omnibus, GSE140021.
Article and author information
Author details
Lilianna Solnica-Krezel
Funding
Children's Discovery Institute
- Lilianna Solnica-Krezel
Washington University School of Medicine in St. Louis (Discretionary funds)
- Lilianna Solnica-Krezel
Vallee Foundation (Vallee Scholar Award)
- Samantha A Morris
Paul G. Allen Frontiers Group (Allen Distinguished Investigator Award)
- Samantha A Morris
Alfred P. Sloan Foundation (Sloan Research Fellowship)
- Samantha A Morris
The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.
Copyright
© 2020, Minn et al.
This article is distributed under the terms of the Creative Commons Attribution License permitting unrestricted use and redistribution provided that the original author and source are credited.
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