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Long-Term Culture of Male Germline Stem Cells From Hamster Testes¹

Department of Molecular Genetics,³ Graduate School of Medicine, Kyoto University, Kyoto 606-8501, Japan Graduate School of Comprehensive Scientific Research,⁴ Prefectural University of Hiroshima, Hiroshima 727-0023, Japan Department of Development and Differentiation,⁵ Institute for Frontier Medical Sciences, Kyoto University, Kyoto 606-8507, Japan

ABSTRACT

Spermatogonial stem cells provide the foundation for spermatogenesis in male animals. We recently succeeded in culturing and genetically engineering mouse spermatogonial stem cells, but little is known regarding the culture and growth requirements of spermatogonial stem cells in other animal species. In this study, we report the successful long-term culture of spermatogonial stem cells from hamster testes. Spermatogonial stem cells were purified using an anti-ITGA6 antibody and cultured in the presence of glial cell line-derived neurotrophic factor. The cells continued to proliferate for at least 1 year. During this period, they were genetically modified using a lentivirus and underwent spermatogenesis after transplantation into the testes of immunodeficient nude mice. However, germ cells generated in the surrogate xenogeneic recipients did not differentiate beyond the spermatid stage, and these round spermatids could not produce offspring through in vitro microinsemination. These results suggest that the germ cells may not have acquired characteristics necessary for fertility in the xenogeneic microenvironment. Nevertheless, the successful establishment of culture conditions conducive for hamster spermatogonial stem cell growth and maintenance indicates that this technique can be extended to other animal species in which current genetic modification techniques are impossible or inefficient.

developmental biology, gametogenesis, spermatogenesis, testis

¹Supported by the Ministry of Education, Culture, Sports, Science, and Technology of Japan, Genome Network Project, Takeda Science Foundation, The Nakajima Foundation, Ichiro Kanehara Foundation, Kowa Life Science, Program for Promotion of Fundamental Studies in Health Sciences of the National Institute of Biomedical Innovation, and Suzuken Memorial Foundation.

Correspondence: ²Takashi Shinohara, Department of Molecular Genetics, Graduate School of Medicine, Kyoto University, Yoshida Konoe, Sakyo-ku, Kyoto 606-8501, Japan. FAX: 81 75 751 4169; e-mail: tshinoha{at}virus.kyoto-u.ac.jp

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