In vitro culture of mouse blastocysts beyond the implantation stages

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

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Blastocyst / cytology*
  • Blastocyst / physiology
  • Cell Adhesion
  • Ectoderm / cytology
  • Embryo Culture Techniques / instrumentation
  • Embryo Culture Techniques / methods*
  • Embryo Implantation
  • Female
  • Mice, Inbred C57BL