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Volume 6, Issue 2 (June 2004) 6, 93–98;
Percoll fractionation of adult mouse spermatogonia improves germ cell transplantation
Kyu-Bom Koh1, Masatoshi Komiyama1, 2, Yoshiro Toyama3, Tetsuya Adachi4, Chisato Mori1, 5
1Department of Bioenvironmental Medicine, 3Department of Anatomy and Developmental Biology, Graduate School of Medicine, Chiba University, Chiba 260-8670, Japan
2Center for Environment, Health and Field Sciences, Chiba University, Kashiwa 277-0882, Japan
4Department of Genomic Drug Discovery Sciences, Graduate School of Pharmaceutical Sciences, Kyoto University, Kyoto 606-8501, Japan
5Core Research for Evolutional Science and Technology, Japan Science and Technology Agency, Kawaguchi 332-0012, Japan
Correspondence: Professor Chisato Mori, E-mail: cmori@faculty.chiba-u.jp
Received 2003-12-24 Accepted 2004-3-23
| Abstract |
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Aim: To isolate and transplant germ cells from adult mouse testes for transplantation. Methods: In order to distinguish transplanted cells from endogenous cells of recipients, donor transgenic mice expressing green fluorescent protein (GFP) were used. Germ cells were collected from the donors at 10-12 weeks of age and spermatogonia were concentrated by percoll fractionation and transplanted into recipient seminiferous tubules that had been previously treated with busulfan at 5 weeks of age to remove the endogenous spermatogenic cells. Results: Twenty weeks after the transplantation, a wide spread GFP signal was observed in the recipient seminiferous tubules. The presence of spermatogenesis and spermatozoa was confirmed in sections of 12 out of 14 testes transplanted (86 %). However, when germ cells were transplanted without concentration the success rate was zero (0/9). Conclusion: Germ cells from adult mouse testes can be successfully transplanted into recipient seminiferous tubules if the cell population is rich in spermatogonia and the percoll fractionation is useful in obtaining such a cell population.
Keywords: germ cell transplantation; green fluorescent protein; percoll fractionation; spermatogenic cells; mice
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