High-resolution transcriptional and morphogenetic profiling of cells from micropatterned human ESC gastruloid cultures

Abstract
Data availability
Article and author information

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.

The following data sets were generated

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2. Fu YC
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4. Dietmann S
5. George SC
6. Anastasio MA
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NCBI Gene Expression Omnibus, GSE144897.

https://www.ncbi.nlm.nih.gov/geo/query/acc.cgi?acc=GSE144897

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Article and author information

Author details

Kyaw Thu Minn

Department of Biomedical Engineering, Washington University in St Louis, St Louis, United States

Competing interests
No competing interests declared.
Yuheng C Fu

Developmental Biology, and Genetics, Washington University School of Medicine, St Louis, United States

Competing interests
No competing interests declared.

"This ORCID iD identifies the author of this article:" 0000-0001-8037-6398
Shenghua He

Department of Computer Science & Engineering, Washington University in St Louis, St Louis, United States

Competing interests
No competing interests declared.
Sabine Dietmann

Department of Developmental Biology, Washington University School of Medicine, St Louis, United States

Competing interests
No competing interests declared.
Steven C George

Department of Biomedical Engineering, University of California, Davis, Davis, United States

Competing interests
No competing interests declared.
Mark A Anastasio

Department of Bioengineering, University of Illinois, Urbana-Champaign, Urbana-Champaign, United States

Competing interests
No competing interests declared.
Samantha A Morris

Developmental Biology, and Genetics, Washington University School of Medicine, St Louis, United States

For correspondence
s.morris@wustl.edu

Competing interests
No competing interests declared.

"This ORCID iD identifies the author of this article:" 0000-0001-8561-4340
Lilianna Solnica-Krezel

Department of Developmental Biology, Washington University School of Medicine, St Louis, United States

For correspondence
solnical@wustl.edu

Competing interests
Lilianna Solnica-Krezel, Reviewing editor, eLife.

"This ORCID iD identifies the author of this article:" 0000-0003-0983-221X

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

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.