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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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Submitted February 13, 2002
Returned for revision March 7, 2002
Accepted August 5, 2002

Testis


Expansion of Murine Spermatogonial Stem Cells Through Serial Transplantation

Takehiko Ogawa 1*, Masako Ohmura 1, Yasushi Yumura 1, Hajime Sawada 1, Yoshinobu Kubota 1
1 Yokohama City University

* To whom correspondence should be addressed. E-mail: ogawatmy{at}mud.biglobe.ne.jp.

Abstract

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 germ cells once established colonies 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 4 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, 1 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.



Key words: Testis • Sperm • Spermatogenesis


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