Mouse Fetal Germ Cell Isolation and Culture Techniques

Methods Mol Biol. 2017:1463:173-183. doi: 10.1007/978-1-4939-4017-2_13.

Abstract

The fetal gonad contains a great variety of differentiating cell populations, of which germ cells make up a small percentage. In order to study germ cell-specific gene and protein expression, as well as determine direct effects of signaling molecules, it is necessary to prepare enriched populations of germ cells and maintain them in culture for several hours to multiple days. The protocols in this chapter are designed to provide a guide for the isolation or enrichment of mouse primordial germ cells (from 9.5 days postcoitum (dpc) to 18.5 dpc) by flow cytometry (Subheading 3.1) or magnetic sorting (Subheading 3.2), followed by primary germ cell culture (Subheading 3.3).

Keywords: FACS; Germ cells; MACS; Magnetic sorting; Serum-free.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cell Culture Techniques / methods*
  • Cell Differentiation
  • Cell Separation / methods*
  • Culture Media, Serum-Free
  • Fetus / cytology*
  • Fetus / metabolism
  • Flow Cytometry
  • Germ Cells / cytology*
  • Germ Cells / metabolism
  • Mice
  • Signal Transduction

Substances

  • Culture Media, Serum-Free