Micropattern differentiation of mouse pluripotent stem cells recapitulates embryo regionalized cell fate patterning

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Abstract

During gastrulation epiblast cells exit pluripotency as they specify and spatially arrange the three germ layers of the embryo. Similarly, human pluripotent stem cells (PSCs) undergo spatially organized fate specification on micropatterned surfaces. Since in vivo validation is not possible for the human, we developed a mouse PSC micropattern system and, with direct comparisons to mouse embryos, reveal the robust specification of distinct regional identities. BMP, WNT, ACTIVIN and FGF directed mouse epiblast-like cells to undergo an epithelial-to-mesenchymal transition and radially pattern posterior mesoderm fates. Conversely, WNT, ACTIVIN and FGF patterned anterior identities, including definitive endoderm. By contrast, epiblast stem cells, a developmentally advanced state, only specified anterior identities, but without patterning. The mouse micropattern system offers a robust scalable method to generate regionalized cell types present in vivo, resolve how signals promote distinct identities and generate patterns, and compare mechanisms operating in vivo and in vitro and across species.

Data availability

The following previously published data sets were used

1. Hayashi
(2011) Reconstitution of the mouse germ-cell specification pathway in culture by pluripotent stem cells
Publicly available at the NCBI Gene Expression Omnibus (accession no: GSE30056).

https://www.ncbi.nlm.nih.gov/geo/query/acc.cgi?acc=GSE30056

Article and author information

Author details

Sophie M Morgani

Developmental Biology Program, Memorial Sloan Kettering Cancer Center, New York, United States

Competing interests
The authors declare that no competing interests exist.
Jakob J Metzger

Center for Studies in Physics and Biology, Rockefeller University, New York, United States

Competing interests
The authors declare that no competing interests exist.
Jennifer Nichols

Wellcome Trust-MRC Center for Stem Cell Research, University of Cambridge, Cambridge, United Kingdom

Competing interests
The authors declare that no competing interests exist.
Eric D Siggia

Center for Studies in Physics and Biology, Rockefeller University, New York, United States

For correspondence
siggiae@mail.rockefeller.edu

Competing interests
The authors declare that no competing interests exist.

"This ORCID iD identifies the author of this article:" 0000-0001-7482-1854
Anna-Katerina Hadjantonakis

Developmental Biology Program, Memorial Sloan Kettering Cancer Center, New York, United States

For correspondence
hadj@mskcc.org

Competing interests
The authors declare that no competing interests exist.

"This ORCID iD identifies the author of this article:" 0000-0002-7580-5124

Funding

National Institute of Diabetes and Digestive and Kidney Diseases (R01DK084391)

Anna-Katerina Hadjantonakis

National Cancer Institute (P30CA008748)

Anna-Katerina Hadjantonakis

Eunice Kennedy Shriver National Institute of Child Health and Human Development (R01HD080699)

Eric D Siggia

National Science Foundation (PHY1502151)

Eric D Siggia

Wellcome

Sophie M Morgani

The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.

Ethics

Animal experimentation: Animal experimentation: All mice used in this study were maintained in accordance withthe guidelines of the Memorial Sloan Kettering Cancer Center (MSKCC) Institutional Animal Care and Use Committee (IACUC) under protocol number 03-12-017 (PI Hadjantonakis).

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.