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Mol. Hum. Reprod. Advance Access published online on June 27, 2009
Molecular Human Reproduction, doi:10.1093/molehr/gap052
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© The Author 2009. Published by Oxford University Press on behalf of the European Society of Human Reproduction and Embryology. All rights reserved. For Permissions, please email: journals.permissions@oxfordjournals.org

New horizons for in vitro spermatogenesis? An update on novel three-dimensional culture systems as tools for meiotic and postmeiotic differentiation of testicular germ cells

Jan-Bernd Stukenborg1,3, Stefan Schlatt1,§, Manuela Simoni1,4, Ching-Hei Yeung1, Mahmoud Abu Elhija2, Craig Marc Luetjens1,5, Mahmoud Huleihel2 and Joachim Wistuba1

1 Institute of Reproductive and Regenerative Biology of the Centre of Reproductive Medicine and Andrology, University Münster, Münster, Germany 2The Shraga Segal Department of Microbiology and Immunology, Faculty of Health Sciences and Soroka University Medical Centre, Ben-Gurion-University of the Negev, Beer-Sheva, Israel

§ Corresponding author: Stefan Schlatt, Institute of Reproductive and Regenerative Biology of the Centre of Reproductive Medicine and Andrology, University Münster, Domagkstrasse 11, 48129 Münster, Germany; Stefan.Schlatt{at}ukmuenster.de

Culture and differentiation of male germ cells has been performed for various purposes in the past. To date, none of the studies aimed at in-vitro spermatogenesis have resulted in a sufficient number of mature gametes. Numerous studies have revealed worthy pieces of information, building up a body of information on conditions that are required to maintain and mature male germ cells in-vitro. In this review, we report on previously published and unpublished experiments addressing murine germ cell differentiation in three-dimensional in-vitro culture systems. In a systematic set of experiments, we examined the influence of two different matrices (soft agar and methylcellulose) as well as the need for gonadotropin support. For the first time, we demonstrate that pre-meiotic male germ cells (revealed by the absence of meiotic marker expression (e.g. Boule)) obtained from immature mice pass through meiosis in vitro. After several weeks of culture, we obtained morphologically normal spermatozoa embedded in the matrix substance. Complete maturation relied on support from somatic testicular cells and the presence of gonadotropins but appeared independent from the matrix in a three-dimensional culture environment. Further research efforts are required to reveal the applicability of this culture technique for human germ cells and the functionality of the spermatozoa for generating offspring.

Key Words: in-vitro spermatogenesis/gonadotropins/SACS

3 present address: Pediatric Endocrinology Unit, Department of Women's and Children's Health, Astrid Lindgren Children's Hospital, Karolinska Institute, Stockholm, Sweden

4 present address: University of Modena and Reggio Emilia, Dept. of Medicine, Endocrinology, Metabolism and Geriatrics, Modena, Italy

5 present address: Study Director, Covance Laboratories, Münster, Germany

Submitted on May 7, 2009; resubmitted on June 12, 2009; accepted on June 19, 2009.


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