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Functional Assessment of Self-Renewal Activity of Male Germline Stem Cells Following Cytotoxic Damage and Serial Transplantation¹

Department of Medical Chemistry,³ Department of Pathology and Biology of Diseases,⁴ Department of Clinical Epidemiology,⁵ Department of Pharmacoepidemiology,⁶ Graduate School of Medicine, Kyoto University, Kyoto 606-8501, Japan

Spermatogenesis is dependent on a small population of stem cells. Although stem cells are believed to expand infinitely, there is little functional evidence regarding whether spermatogonial stem cells can increase in their number. Using the spermatogonial transplantation technique, we evaluated the proliferative potential of spermatogonial stem cells in two models of regeneration. After busulfan injection to deplete stem cells, the surviving stem cells were able to expand by at least 15.8-fold within 2 mo. On the other hand, a serial transplantation study indicated that one transplanted stem cell was able to expand by 3.8- and 12-fold within 2 and 4 mo, respectively. These results provide direct functional evidence for the expansion of stem cells and establish the basis for further characterization of the stem cell self-renewal process.

¹ M.K.-S. was supported by a grant from the Japan Society for the Promotion of Science. Financial support for this research was provided by the Kanae Foundation for Life and Socio-Medical Science; the Inamori Foundation; and the Ministry of Education, Science, Sports, and Culture of Japan.

² Correspondence: Takashi Shinohara, Department of Medical Chemistry, Graduate School of Medicine, Kyoto University, Yoshida-Konoe, Sakyo-ku, Kyoto 606-8501, Japan. FAX: 81 75 753 4388; takashi{at}mfour.med.kyoto-u.ac.jp

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