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Biological Activity and Enrichment of Spermatogonial Stem Cells in Vitamin A-Deficient and Hyperthermia-Exposed Testes from Mice Based on Colonization Following Germ Cell Transplantation¹

^a School of Molecular Biosciences, Center for Reproductive Biology, Washington State University, Pullman, Washington 99164-4660 ^b Department of Physiology, School of Medicine, Southern Illinois University, Carbondale, Illinois 62901

Spermatogenesis is a complex process in which spermatogonial stem cells divide and subsequently differentiate into spermatozoa. This process requires spermatogonial stem cells to self-renew and provide a continual population of cells for differentiation. Studies on spermatogonial stem cells have been limited due to a lack of unique markers and an inability to detect the presence of these cells. The technique of germ cell transplantation provides a functional assay to identify spermatogonial stem cells in a cell population. We hypothesized that vitamin A-deficient (VAD) and hyperthermically treated testes would provide an enriched in vivo source of spermatogonial stem cells. The first model, hyperthermic treatment, depends on the sensitivity of maturing germ cells to high temperatures. Testes of adult mice were exposed to 43°C for 15 min to eliminate the majority of differentiating germ cells. Treated donor testes were 50% of normal adult testis size and, when transplanted into recipients, resulted in a 5.3- and 19-fold (colonies and area, respectively) increase in colonization efficiency compared to controls. The second model, VAD animals, also lacked differentiating germ cells, and testes weights were 25% of control values. Colonization efficiency of germ cells from VAD testes resulted in a 2.5- and 6.2-fold (colonies and area, respectively) increase in colonization compared to controls. Hyperthermically treated mice represent an enriched source of spermatogonial stem cells. In contrast, the low extent of colonization with germ cells from VAD animals raises important questions regarding the competency of stem cells from this model.

First decision: 28 August 2001.

¹ Supported by NIH grant (National Institute of Child Health and Human Development) HD 35494 to M.D.G. and HD 08577-02 to D.J.M.

² Correspondence: Michael D. Griswold, School of Molecular Biosciences, Washington State University, Box 644660, Pullman, WA 99164-4660. FAX: 509 335 9688; griswold{at}mail.wsu.edu

³ Deceased

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