HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS		QUICK SEARCH:   [advanced] Author: Keyword(s): Year:  Vol:  Page:

This Article Full Text Full Text (PDF) All Versions of this Article: 71/3/942    most recent biolreprod.104.028894v1 Alert me when this article is cited Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal My Folders Download to citation manager Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Oatley, J. M. Articles by McLean, D. J. Search for Related Content PubMed PubMed Citation Articles by Oatley, J. M. Articles by McLean, D. J. Agricola Articles by Oatley, J. M. Articles by McLean, D. J.

Biological Activity of Cryopreserved Bovine Spermatogonial Stem Cells During In Vitro Culture¹

Department of Animal Sciences and Center for Reproductive Biology, Washington State University, Pullman, Washington 99164

Functional roles of spermatogonial stem cells in spermatogenesis are self-renewing proliferation and production of differentiated daughter progeny. The ability to recapitulate these actions in vitro is important for investigating their biology and inducing genetic modification that could potentially lead to an alternative means of generating transgenic animals. The objective of this study was to evaluate the survival and proliferation of frozen-thawed bovine spermatogonial stem cells in vitro and investigate the effects of exogenous glial cell line-derived neurotrophic factor (GDNF). In order to accomplish this objective we developed a bovine embryonic fibroblast feeder cell line, termed BEF, to serve as feeder cells in a coculture system with bovine germ cells. Bovine spermatogonial stem cell survival and proliferation in vitro were evaluated by xenogeneic transplantation into the seminiferous tubules of immunodeficient mice. Bovine germ cells cocultured for 1 wk resulted in significantly more round cell donor colonies in recipient mouse testes compared to donor cells transplanted just after thawing. Bovine germ cells cocultured for 2 wk had fewer colony-forming cells than the freshly thawed cell suspensions or cells cultured for 1 wk. Characterization of the feeder cell line revealed endogenous expression of Gdnf mRNA and protein. Addition of exogenous GDNF to the culture medium decreased the number of stem cells present at 1 wk of coculture, but enhanced stem cell maintenance at 2 wk compared to cultures without added GDNF. These data indicate that frozen-thawed bovine spermatogonial stem cells survive cryopreservation and can be maintained during coculture with a feeder cell line in which the maintenance is influenced by GDNF.

¹ J.M.O. was supported by an Achievement Rewards for Collegiate Scientists (ARCS) Fellowship. J.J.R. was supported by a Baxter Endowed Grant from Washington State University.

² Correspondence. FAX: 509 335 4246; dmclean{at}wsu.edu

This article has been cited by other articles:

				J. A. Schmidt, M. R. Avarbock, J. W. Tobias, and R. L. Brinster Identification of Glial Cell Line-Derived Neurotrophic Factor-Regulated Genes Important for Spermatogonial Stem Cell Self-Renewal in the Rat Biol Reprod, July 1, 2009; 81(1): 56 - 66. [Abstract] [Full Text] [PDF]

				C. B. Maki, J. Pacchiarotti, T. Ramos, M. Pascual, J. Pham, J. Kinjo, S. Anorve, and F. Izadyar Phenotypic and molecular characterization of spermatogonial stem cells in adult primate testes Hum. Reprod., June 1, 2009; 24(6): 1480 - 1491. [Abstract] [Full Text] [PDF]

				P. M Aponte, T. Soda, K. J Teerds, S C. Mizrak, H. J G van de Kant, and D. G de Rooij Propagation of bovine spermatogonial stem cells in vitro Reproduction, November 1, 2008; 136(5): 543 - 557. [Abstract] [Full Text] [PDF]

				J. A. Schmidt, J. M. d. Avila, and D. J. McLean Analysis of Gene Expression in Bovine Testis Tissue Prior to Ectopic Testis Tissue Xenografting and During the Grafting Period Biol Reprod, June 1, 2007; 76(6): 1071 - 1080. [Abstract] [Full Text] [PDF]

				S. Fouchecourt, M. Godet, O. Sabido, and P. Durand Glial cell-line-derived neurotropic factor and its receptors are expressed by germinal and somatic cells of the rat testis. J. Endocrinol., July 1, 2006; 190(1): 59 - 71. [Abstract] [Full Text] [PDF]

				J. M. Oatley, M. R. Avarbock, A. I. Telaranta, D. T. Fearon, and R. L. Brinster From the Cover: Identifying genes important for spermatogonial stem cell self-renewal and survival PNAS, June 20, 2006; 103(25): 9524 - 9529. [Abstract] [Full Text] [PDF]

				M. Kanatsu-Shinohara, K. Inoue, J. Lee, H. Miki, N. Ogonuki, S. Toyokuni, A. Ogura, and T. Shinohara Anchorage-Independent Growth of Mouse Male Germline Stem Cells In Vitro Biol Reprod, March 1, 2006; 74(3): 522 - 529. [Abstract] [Full Text] [PDF]