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In vitro generation of mouse polarized embryo-like structures from embryonic and trophoblast stem cells

Harrison, Sarah Ellys and Sozen, Berna and Zernicka-Goetz, Magdalena (2018) In vitro generation of mouse polarized embryo-like structures from embryonic and trophoblast stem cells. Nature Protocols, 13 (7). pp. 1586-1602. ISSN 1754-2189. doi:10.1038/s41596-018-0005-x. https://resolver.caltech.edu/CaltechAUTHORS:20190405-170314258

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Abstract

Mammalian embryogenesis requires the coordination of embryonic and extra-embryonic tissues to enable implantation into the uterus and post-implantation development to establish the body plan. Mouse embryonic stem cells (ESCs) are a useful tool for studying pluripotent embryonic tissue in vitro. However, they cannot undertake correct embryogenesis alone. Many attempts to model the early embryo in vitro involve the aggregation of ESCs into spheroids of variable size and cell number that undertake germ-layer specification but fail to recapitulate the characteristic architecture and arrangement of tissues of the early embryo. Here, we describe a protocol to generate the first embryo-like structures by directing the assembly of mouse ESCs and extra-embryonic trophoblast stem cells (TSCs) in a 3D extracellular matrix (ECM) into structures we call ‘polarized embryo-like structures’. By establishing the medium and culture conditions needed to support the growth of both stem cell types simultaneously, embryonic architecture is generated within 4 d of co-culture. This protocol can be performed by those proficient in standard ESC culture techniques and can be used in developmental studies to investigate the interactions between embryonic and extra-embryonic tissues during mammalian development.


Item Type:Article
Related URLs:
URLURL TypeDescription
https://doi.org/10.1038/s41596-018-0005-xDOIArticle
https://rdcu.be/bvvEgPublisherFree ReadCube access
ORCID:
AuthorORCID
Harrison, Sarah Ellys0000-0002-1571-2399
Sozen, Berna0000-0001-5834-5819
Zernicka-Goetz, Magdalena0000-0002-7004-2471
Additional Information:© 2018 Springer Nature Publishing AG. Published online: 9 July 2018. We are grateful to our colleagues in the M.Z.-G. group and to M. Huch for advice. We are grateful to the Wellcome Trust for the Senior Research fellowship (grant no. 098287/Z/12/Z) and for a European Research Council grant (code: 669198) awarded to M.Z.-G. to fund this work. We are also grateful for the BBSRC DTP studentship that supports S.E.H. and to the Scientific and Technological Research Council of Turkey, which supports B.S.’ These authors contributed equally: Sarah Ellys Harrison and Berna Sozen. Author Contributions: S.E.H., B.S., and M.Z.-G. designed the protocol, carried out the work, analyzed the results, and prepared the manuscript. Competing interests: M.Z.-G. and S.E.H. declare that they are inventors on a patent application (1615343.9) submitted by Cell Guidance Systems (in which the University of Cambridge and the Wellcome Trust are beneficiaries) that covers the method and medium composition used to generate stem cell–derived embryos. B.S. declares no competing interests.
Funders:
Funding AgencyGrant Number
Wellcome Trust098287/Z/12/Z
European Research Council (ERC)669198
Biotechnology and Biological Sciences Research Council (BBSRC)UNSPECIFIED
Türkiye Bilimsel ve Teknolojik Araştırma Kurumu (TÜBİTAK) UNSPECIFIED
Issue or Number:7
DOI:10.1038/s41596-018-0005-x
Record Number:CaltechAUTHORS:20190405-170314258
Persistent URL:https://resolver.caltech.edu/CaltechAUTHORS:20190405-170314258
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:94529
Collection:CaltechAUTHORS
Deposited By: George Porter
Deposited On:09 Apr 2019 14:55
Last Modified:16 Nov 2021 17:05

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