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In vitro culture of mouse blastocysts beyond the implantation stages

Bedzhov, Ivan and Leung, Chuen Yan and Bialecka, Monika and Zernicka-Goetz, Magdalena (2014) In vitro culture of mouse blastocysts beyond the implantation stages. Nature Protocols, 9 (12). pp. 2732-2739. ISSN 1754-2189. doi:10.1038/nprot.2014.186.

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The implanting mouse blastocyst invades the uterine stroma and undergoes a dramatic transformation into an egg cylinder. The morphogenetic and signaling events during this transition are largely unexplored, as the uterine tissues engulf the embryo. Here we describe a protocol supporting the development of the mouse embryo beyond the blastocyst stage in vitro. We established two types of medium to be applied sequentially, and we used a substrate permitting high-resolution imaging of the transition from blastocyst to egg cylinder. We developed two variants of this protocol: the first starts with intact early blastocysts that upon zona removal can attach to the substrate and develop into egg cylinders after 5 d, and the second starts with late blastocysts that upon dissection of the mural trophectoderm form egg cylinders in only 3 d. This method allows observation of a previously hidden period of development, and it provides a platform for novel research into peri-implantation embryogenesis and beyond.

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Zernicka-Goetz, Magdalena0000-0002-7004-2471
Additional Information:© 2014 Nature Publishing Group. Published 30 October 2014. We are grateful to the Wellcome Trust for the Senior Research Fellowship to M.Z.-G. that funded this work. We also thank the European Molecular Biology Organization (EMBO) for supporting M.B. Ivan Bedzhov, Chuen Yan Leung & Monika Bialecka - These authors contributed equally to this work. Author Contributions: I.B., C.Y.L., M.B. and M.Z.-G. designed the protocol, carried out the work, analyzed the results and prepared the manuscript. The authors declare no competing financial interests.
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European Molecular Biology Organization (EMBO)UNSPECIFIED
Issue or Number:12
Record Number:CaltechAUTHORS:20190405-170316220
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Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:94548
Deposited By: George Porter
Deposited On:10 Apr 2019 14:59
Last Modified:16 Nov 2021 17:05

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