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).
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Szabo PE, Hubner K, Scholer H, Mann JR (2002) Allele-specific expression of imprinted genes in mouse migratory primordial germ cells. Mech Dev 115(1–2):157–160, doi:S0925477302000874 [pii]
Durcova-Hills G, Surani A (2008) Reprogramming primordial germ cells (PGC) to embryonic germ (EG) cells. Curr Protoc Stem Cell Biol Chapter 1:Unit1A 3. doi:10.1002/9780470151808.sc01a03s5
Solter D, Knowles BB (1978) Monoclonal antibody defining a stage-specific mouse embryonic antigen (SSEA-1). Proc Natl Acad Sci U S A 75(11):5565–5569
Donovan PJ, Stott D, Godin I, Heasman J, Wylie CC (1987) Studies on the migration of mouse germ cells. J Cell Sci Suppl 8:359–367
Mackay S, Nicholson CL, Lewis SP, Brittan M (1999) E-cadherin in the developing mouse gonad. Anat Embryol (Berl) 200(1):91–102
Farini D, Scaldaferri ML, Iona S, La Sala G, De Felici M (2005) Growth factors sustain primordial germ cell survival, proliferation and entering into meiosis in the absence of somatic cells. Dev Biol 285(1):49–56. doi:10.1016/j.ydbio.2005.06.036
De Miguel MP, Donovan PJ (2000) Isolation and culture of mouse germ cells. Methods Mol Biol 137:403–408. doi:10.1385/1-59259-066-7:403
Resnick JL, Bixler LS, Cheng L, Donovan PJ (1992) Long-term proliferation of mouse primordial germ cells in culture. Nature 359(6395):550–551. doi:10.1038/359550a0
Matsui Y, Zsebo K, Hogan BL (1992) Derivation of pluripotential embryonic stem cells from murine primordial germ cells in culture. Cell 70(5):841–847, doi:0092-8674(92)90317-6 [pii]
Donovan J, Brown P (2006) Euthanasia. Curr Protoc Immunol Chapter 1:Unit 1 8. doi:10.1002/0471142735.im0108s73
McFarlane L, Truong V, Palmer JS, Wilhelm D (2013) Novel PCR assay for determining the genetic sex of mice. Sex Dev 7(4):207–211. doi:10.1159/000348677
Lawson KA, Dunn NR, Roelen BA, Zeinstra LM, Davis AM, Wright CV, Korving JP, Hogan BL (1999) Bmp4 is required for the generation of primordial germ cells in the mouse embryo. Genes Dev 13(4):424–436
Acknowledgments
This work was supported by the Australian Research Council [DP140104059] and Cancer Council Queensland [APP1012325]. C.S. is a University of Queensland Postdoctoral Fellow.
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2017 Springer Science+Business Media New York
About this protocol
Cite this protocol
Spiller, C.M., Burnet, G., Bowles, J. (2017). Mouse Fetal Germ Cell Isolation and Culture Techniques. In: Buszczak, M. (eds) Germline Stem Cells. Methods in Molecular Biology, vol 1463. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-4017-2_13
Download citation
DOI: https://doi.org/10.1007/978-1-4939-4017-2_13
Published:
Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-4015-8
Online ISBN: 978-1-4939-4017-2
eBook Packages: Springer Protocols