Journal of Andrology
HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH
Published-Ahead-of-Print November 28, 2007, DOI:10.2164/jandrol.107.002857

This Article
Right arrow Author Manuscript (PDF)
Right arrow All Versions of this Article:
29/3/312    most recent
Author Manuscript (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Google Scholar
Right arrow Articles by Stukenborg, J.-B.
Right arrow Articles by Schlatt, S.
PubMed
Right arrow PubMed Citation
Right arrow Articles by Stukenborg, J.-B.
Right arrow Articles by Schlatt, S.

Co-culture of spermatogonia with somatic cells in a novel three-dimensional Soft-Agar-Culture-System

Jan-Bernd Stukenborg , Joachim Wistuba , Craig Marc Luetjens , Mahmoud Abu Elhija , Mahmoud Huleihel , Eitan Lunenfeld , Jörg Gromoll , Eberhard Nieschlag *, and Stefan Schlatt

* To whom correspondence should be addressed. E-mail: eberhard.nieschlag{at}ukmuenster.de.

Isolation and culture of spermatogonial stem cells (SSCs) has become an approach to study the milieu and the factors controlling their expansion and differentiation. Traditional conventional cell culture does not mimic the complex situation in the seminiferous epithelium providing a basal, intraepithelial, and adluminal compartment to the developing male germ cells. SSCs are located in specific stem cell niches whose features and functional parameters are thus far poorly understood. It was the aim of this study to isolate SSCs and to explore their expansion and differentiation potential in a novel three-dimensional Soft-Agar-Culture-System (SACS). This system provides three-dimensional structural support and multiple options for manipulations through the addition of factors, cells or other changes. The system has revolutionized research on blood stem cells by providing a tool for clonal analysis of expanding and differentiating blood cell lineages. In our studies SSCs are enriched using Gfr{alpha}-1 as a specific surface marker and magnetic activated cell sorting (MACS) as a separation approach. At termination of the culture we determined the type and number of germ cells obtained after the first 24 hours of culture. We also determined cell types and numbers in expanding cell clones of differentiating germ cells during the subsequent 15 days of culture. We analyzed a supportive effect of somatic cell lineages added to the solid part of the culture system. We conclude that our enrichment and culture approach is highly useful for exploration of SSC expansion and have found indications that the system supports differentiation up to the level of postmeiotic germ cells.


Key words: Spermatogenesis • Testis • Soft-Agar-Culture-System • in vitro meiosis • spermatogonial stem cells






HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH
Copyright © 2007 by The American Society of Andrology.