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Nat Protoc. 2014 Dec;9(12):2732-9. doi: 10.1038/nprot.2014.186. Epub 2014 Oct 30.

In vitro culture of mouse blastocysts beyond the implantation stages.

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1
1] Department of Physiology, Development and Neuroscience, Downing Site, University of Cambridge, Cambridge, UK. [2] Wellcome Trust/Cancer Research UK Gurdon Institute, University of Cambridge, Cambridge, UK.

Abstract

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.

PMID:
25356584
DOI:
10.1038/nprot.2014.186
[Indexed for MEDLINE]
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