Isolation and Culture Techniques for Fetal Mouse Germ Cells

Guillaume Burnet; Josephine Bowles; Cassy M Spiller

doi:10.1007/978-1-0716-3259-8_13

Isolation and Culture Techniques for Fetal Mouse Germ Cells

Methods Mol Biol. 2023:2677:221-231. doi: 10.1007/978-1-0716-3259-8_13.

Authors

Guillaume Burnet¹, Josephine Bowles^{1

2}, Cassy M Spiller³

Affiliations

¹ School of Biomedical Sciences, The University of Queensland, Brisbane, QLD, Australia.
² Institute for Molecular Bioscience, The University of Queensland, Brisbane, QLD, Australia.
³ School of Biomedical Sciences, The University of Queensland, Brisbane, QLD, Australia. c.spiller@uq.edu.au.

PMID: 37464245
DOI: 10.1007/978-1-0716-3259-8_13

Abstract

The fetal gonad contains a great variety of differentiating cell populations, of which germ cells make up a relatively 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 primordial germ cells (from 9.5 days post coitum (dpc) to 18.5 dpc) by flow cytometry (Subheading 3.1) or magnetic sorting (Subheading 3.2), followed by feeder-free 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
Culture Techniques
Fetus*
Flow Cytometry
Germ Cells* / metabolism
Gonads
Mice