Abstract
The survival, proliferation, and differentiation of primordial germ cells in the mammalian embryo is regulated by a complex cocktail of growth factors and interactions with surrounding somatic cells, which together form a microenvironment known as the germ cell niche. Extensive insight into the signalling pathways that regulate PGC behaviour has been provided by the study of these cells in rodent models, however little is known about the factors that regulate these processes in human PGCs. In this review, we outline experimental approaches to the culture and manipulation of the first trimester human fetal ovary, and discuss immunohistochemical and stereological approaches to detect changes in human PGC numbers and proliferation in response to treatment with exogenous growth factors.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Matzuk MM, Lamb DJ (2008) The biology of infertility: research advances and clinical challenges. Nat Med 14:1197–1213.
Anderson RA, Fulton N, Cowan G, Coutts S, Saunders PT (2007) Conserved and divergent patterns of expression of DAZL, VASA and OCT4 in the germ cells of the human fatal ovary and testis. BMC Dev Biol 7:136.
Witschi E (1948) Migration of the germ cells of human embryos from the yolk sac to the primitive gonadal folds Contrib Embryol 32:67–80.
Hanley NA, Hagan DM, Clement-Jones M, Ball SG, Strachan T, Salas-Cortes L, McElreavey K, Lindsay S, Robson S, Bullen P, Ostrer H, Wilson DI (2000) SRY, SOX9, and DAX1 expression patterns during human sex determination and gonadal development. Mech Dev 91:403–407.
Wartenburg H (1981) Differentiation and development of the testes, in The Testis (Burger, H., de Kretser, D.M., Ed.), 2nd ed., Raven Press, New York.
Bendsen E, Byskov AG, Andersen CY, Westergaard LG (2006) Number of germ cells and somatic cells in human fatal ovaries during the first weeks after sex differentiation. Hum Reprod 21:30–35.
Gondos B, Westergaard L, Byskov AG (1986) Initiation of oogenesis in the human fatal ovary: ultrastructural and squash preparation study. Am J Obstet Gynecol 155:189–195.
Le Bouffant R, Guerquin MJ, Duquenne C, Frydman N, Coffigny H, Rouiller-Fabre V, Frydman R, Habert R, Livera G (2010) Meiosis initiation in the human ovary requires intrinsic retinoic acid synthesis. Hum Reprod 25:2579–2590.
Bullejos M, Koopman P (2004) Germ cells enter meiosis in a rostro-caudal wave during development of the mouse ovary. Mol Reprod Dev 68:422–428.
Menke DB, Koubova J, Page DC (2003) Sexual differentiation of germ cells in XX mouse gonads occurs in an anterior-to-posterior wave. Dev Biol 262:303–312.
Anderson G, Gordon K (1996) Tissue processing, microtomy and paraffin sections, in Theory and Practice of Histological Techniques (Bankroft, J. D., and Stevens, A., Eds.) 4th ed., Churchill Livingstone, New York.
Fulton N, Martins da Silva SJ, Bayne RA, Anderson RA (2005) Germ cell proliferation and apoptosis in the developing human ovary. J Clin Endocrinol Metab 90:4664–4670.
Stoop H, Honecker F, Cools M, de Krijger R, Bokemeyer C, Looijenga LH (2005) Differentiation and development of human female germ cells during prenatal gonadogenesis: an immunohistochemical study. Hum Reprod 20:1466–1476.
De Felici M, Scaldaferri ML, Lobascio M, Iona S, Nazzicone V, Klinger FG, Farini D (2004) Experimental approaches to the study of primordial germ cell lineage and proliferation. Hum Reprod Update 10:197–206.
Donovan PJ (1994) Growth factor regulation of mouse primordial germ cell development. Curr Top Dev Biol 29:189–225.
Childs AJ, Kinnell,HL, Collins CS, Hogg K, Bayne RA, Green SJ, McNeilly AS, Anderson RA (2010) BMP signaling in the human fatal ovary is developmentally regulated and promotes primordial germ cell apoptosis. Stem Cells 28:1368–1378.
Childs AJ, Saunders PT, Anderson RA (2008) Modelling germ cell development in vitro. Mol Hum Reprod 14:501–511.
Shamblott MJ, Axelman J, Wang S, Bugg EM, Littlefield JW, Donovan PJ, Blumenthal PD Huggins GR, Gearhart JD (1998) Derivation of pluripotent stem cells from cultured human primordial germ cell., Proc Natl Acad Sci USA 95:13726–13731.
Turnpenny L, Brickwood S, Spalluto CM, Piper K, Cameron IT, Wilson DI, Hanley NA (2003) Derivation of human embryonic germ cells: an alternative source of pluripotent stem cells. Stem Cells 21:598–609.
Lambrot R, Coffigny H, Pairault C, Donnadieu AC, Frydman R, Habert R, Rouiller-Fabre V (2006) Use of organ culture to study the human fatal testis development: effect of retinoic acid. J Clin Endocrinol Metab 91:2696–2703.
Hoei-Hansen CE, Nielsen JE, Almstrup K, Sonne SB, Graem N, Skakkebaek NE, Leffers H, Rajpert-De Meyts E (2004) Transcription factor AP-2gamma is a developmentally regulated marker of testicular carcinoma in situ and germ cell tumors. Clin Cancer Res 10:8521–8530.
Pauls K, Jager R, Weber S, Wardelmann E, Koch A, Buttner R, Schorle H (2005) Transcription factor AP-2gamma, a novel marker of gonocytes and seminomatous germ cell tumors. Int J Cancer 115:470–477.
Bayne RA, Eddie SL, Collins CS, Childs AJ, Jabbour HN, Anderson RA (2009) Prostaglandin E2 as a regulator of germ cells during ovarian development. J Clin Endocrinol Metab 94:4053–4060.
Childs AJ, Bayne RA, Murray AA, Martins Da Silva SJ, Collins CS, Spears N, Anderson RA (2010) Differential expression and regulation by activin of the neurotrophins BDNF and NT4 during human and mouse ovarian development. Dev Dyn 239:1211–1219.
Farhi J, Fisch B, Garor R, Peled Y, Pinkas H, Abir R (2010) Neurotrophin 4 enhances in vitro follicular assembly in human fatal ovaries, Fertil Steril. doi: doi:10.1016/j.fertnstert.2010.03.051.
Friel A, Houghton JA, Glennon M, Lavery R, Smith T, Nolan A, Maher M (2002) A preliminary report on the implication of RT-PCR detection of DAZ, RBMY1, USP9Y and Protamine-2 mRNA in testicular biopsy samples from azoospermic men. Int J Androl 25:59–64.
McClellan KA, Gosden R, Taketo T (2003) Continuous loss of oocytes throughout meiotic prophase in the normal mouse ovary. Dev Biol 258:334–348.
Acknowledgements
We are grateful to Hazel Kinnell, Dr Rosemary Bayne, Sharon Eddie, and the staff of the Medical Research Council Human Reproductive Sciences Unit Histology Core Facility for technical assistance in developing these protocols, and to Anne Saunderson, Joan Creiger, and the staff of the Bruntsfield Suite of the Royal Infirmary of Edinburgh for assistance with patient recruitment and specimen collection. This work is supported by Medical Research Council core funding to RAA (U.1276.00.002.00001.01) and a Medical Research Scotland grant (354 FRG) to AJC.
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2012 Springer Science+Business Media, LLC
About this protocol
Cite this protocol
Childs, A.J., Anderson, R.A. (2012). Experimental Approaches to the Study of Human Primordial Germ Cells. In: Chan, WY., Blomberg, L. (eds) Germline Development. Methods in Molecular Biology, vol 825. Springer, New York, NY. https://doi.org/10.1007/978-1-61779-436-0_15
Download citation
DOI: https://doi.org/10.1007/978-1-61779-436-0_15
Published:
Publisher Name: Springer, New York, NY
Print ISBN: 978-1-61779-435-3
Online ISBN: 978-1-61779-436-0
eBook Packages: Springer Protocols