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BOR - Papers in Press, published online ahead of print April 16, 2003.
Biol Reprod 2003, 10.1095/biolreprod.103.017012
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BIOLOGY OF REPRODUCTION 69, 612–616 (2003)
DOI: 10.1095/biolreprod.103.017012
© 2003 by the Society for the Study of Reproduction, Inc.

Testis

Long-Term Proliferation in Culture and Germline Transmission of Mouse Male Germline Stem Cells1

Mito Kanatsu-Shinohara2,3, Narumi Ogonuki4, Kimiko Inoue5, Hiromi Miki5, Atsuo Ogura5, Shinya Toyokuni4, and Takashi Shinohara3

Horizontal Medical Research Organization,3 Department of Pathology and Biology of Diseases,4 Graduate School of Medicine, Kyoto University, Kyoto 606-8501, Japan The Institute of Physical and Chemical Research (RIKEN),5 Bioresource Center, Ibaraki 305-0074, Japan

Spermatogenesis is a complex process that originates in a small population of spermatogonial stem cells. Here we report the in vitro culture of spermatogonial stem cells that proliferate for long periods of time. In the presence of glial cell line-derived neurotrophic factor, epidermal growth factor, basic fibroblast growth factor, and leukemia inhibitory factor, gonocytes isolated from neonatal mouse testis proliferated over a 5-month period (>1014-fold) and restored fertility to congenitally infertile recipient mice following transplantation into seminiferous tubules. Long-term spermatogonial stem cell culture will be useful for studying spermatogenesis mechanism and has important implications for developing new technology in transgenesis or medicine.

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

2 Correspondence: Mito Kanatsu-Shinohara, Horizontal Medical Research Organization, Graduate School of Medicine, Kyoto University, Yoshida-Konoe, Sakyo-ku, Kyoto 606-8501, Japan. FAX: 81 75 753 9281; mitos{at}mfour.med.kyoto-u.ac.jp




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