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Production of Functional Spermatids from Mouse Germline Stem Cells in Ectopically Reconstituted Seminiferous Tubules¹

Departments of Urology,³ Molecular Pathology,⁴ and Regenerative Medicine,⁵ Yokohama City University Graduate School of Medicine, Yokohama 236-0004, Japan Mitsubishi Kagaku Institute of Life Science,⁶ Tokyo 194-8511, Japan RIKEN,⁷ Bioresource Center, Ibaraki 305-0074, Japan Department of Science for Laboratory Animal Experimentation,⁸ Research Institute for Microbial Diseases, Osaka University, Osaka 565-0871, Japan

ABSTRACT

Testicular germ cell transplantation into the seminiferous tubules is at present the only way to induce spermatogenesis from a given source of spermatogonial stem cells. Here we show an alternative method that harnesses the self-organizing ability of testicular somatic cells. The testicular cells of embryonic or neonatal mice or rats and of newborn pigs were dissociated into single cells. Each of them reorganized into a tubular structure following implantation into the subcutis of immunodeficient mice. When mouse germline stem (GS) cells derived from spermatogonial stem cells and expanded in culture were intermingled with testicular cells of rodents, they were integrated in the reconstituted tubules and differentiated beyond meiosis into spermatids. Normal offspring were produced by the microinjection of those spermatids into oocytes. This method could be applicable to various mammalian species and useful for producing functional gametes from GS cells in a xenoectopic environment.

assisted reproductive technology, gametogenesis, meiosis, spermatogenesis, spermatogonial stem cell, testis

¹Supported in part by the 2005 Strategic Research Project grant K17002 from Yokohama City University, Japan.

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

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