Isolation and manipulation of mouse trophoblast stem cells

Curr Protoc Stem Cell Biol. 2015 Feb 2:32:1E.4.1-1E.4.32. doi: 10.1002/9780470151808.sc01e04s32.

Abstract

The isolation of stable trophoblast stem (TS) cell lines from early mouse embryos has provided a useful cell culture model to study trophoblast development. TS cells are derived from pre-implantation blastocysts or from the extraembryonic ectoderm of early post-implantation embryos. The derivation and maintenance of mouse TS cells is dependent upon continuous fibroblast growth factor (FGF) signaling. Gene expression analysis, differentiation in culture, and chimera formation show that TS cells accurately model the mouse trophoblast lineage. This unit describes how to derive, maintain, and manipulate TS cells, including DNA transfection and chimera formation.

Keywords: FGF4; TS cells; extraembryonic ectoderm; trophectoderm; trophoblast; trophoblast stem cells.

MeSH terms

  • Animals
  • Blastocyst / cytology
  • Cell Count
  • Cell Separation / methods*
  • Cells, Cultured
  • Chimera
  • Coculture Techniques
  • Culture Media, Conditioned / pharmacology
  • DNA / metabolism
  • Electroporation
  • Fibroblasts / cytology
  • Fibroblasts / drug effects
  • Freezing
  • Lipids
  • Mice
  • Mitomycin / pharmacology
  • Stem Cells / cytology*
  • Stem Cells / drug effects
  • Transfection
  • Trophoblasts / cytology*

Substances

  • Culture Media, Conditioned
  • Lipids
  • Lipofectamine
  • Mitomycin
  • DNA