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BOR - Papers in Press, published online ahead of print October 14, 2002.
Biol Reprod 2002, 10.1095/biolreprod.102.004549
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BIOLOGY OF REPRODUCTION 68, 316–322 (2003)
DOI: 10.1095/biolreprod.102.004549
© 2003 by the Society for the Study of Reproduction, Inc.


Testis

Expansion of Murine Spermatogonial Stem Cells Through Serial Transplantation1

Takehiko Ogawa2,a, Masako Ohmurab, Yasushi Yumuraa, Hajime Sawadab, and Yoshinobu Kubotaa

a Department of Urology b Anatomy, Yokohama City University School of Medicine, Yokohama 236-0004, Japan

Mammalian male germ cells might be generally thought to have infinite proliferative potential based on their life-long production of huge numbers of sperm. However, there has been little substantial evidence that supports this assumption. In the present study, we performed serial transplantation of spermatogonial stem cells to investigate if they expand by self-renewing division following transplantation. The transgenic mouse carrying the Green fluorescent protein gene was used as the donor cell source that facilitated identification and recollection of colonized donor germ cells in the recipient testes. The established colonies of germ cells in the recipient testes were collected and transplanted to new recipients. This serial transplantation of spermatogonial stem cells repopulated the recipient testes, which were successfully performed sequentially up to four times from one recipient to the next. The incubation periods between two sequential transplantations ranged from 55 to 373 days. During these passages, the spermatogonial stem cells showed constant activity to form spermatogenic colonies in the recipient testis. They continued to increase in number for more than a year following transplantation. Colonization efficiency of spermatogonial stem cells was determined to be 4.25% by using Sl/Sld mice as recipients that propagated only undifferentiated type A spermatogonia in their testes. Based on the colonization efficiency, one colony-forming activity was assessed to equate to about 20 spermatogonial stem cells. The spermatogonial stem cells were estimated to expand over 50-fold in 100 days in this experiment.

1 This research was supported by a Grant-in-Aid for Scientific Research from the Ministry of Education, Science, Sports, and Culture, Japan (T.O., 11671569), and a Grant-in-Aid from Suzuki Foundation for Urological Medicine (T.O.).

2 Correspondence: T. Ogawa, Department of Urology, Yokohama City University School of Medicine, 3-9 Fukuura, Kanazawa-ku, Yokohama, 236-0004, Japan. FAX: 81 45 786 5775; ogawa{at}med.yokohama-cu.ac.jp




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