title: In vitro culture of mouse blastocysts beyond the implantation stages.
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.
authors.affiliation: 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.
authors.forename: Ivan
authors.initials: I
authors.lastname: Bedzhov
authors.affiliation: 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.
authors.forename: Chuen Yan
authors.initials: CY
authors.lastname: Leung
authors.affiliation: 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.
authors.forename: Monika
authors.initials: M
authors.lastname: Bialecka
authors.affiliation: 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.
authors.forename: Magdalena
authors.initials: M
authors.lastname: Zernicka-Goetz
mesh_terms: D000818:Animals; D001755:Blastocyst / Q000166:cytology* / Q000502:physiology; D002448:Cell Adhesion; D004475:Ectoderm / Q000166:cytology; D046149:Embryo Culture Techniques / Q000295:instrumentation / Q000379:methods*; D010064:Embryo Implantation; D005260:Female; D008810:Mice, Inbred C57BL