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Recipient Preparation and Mixed Germ Cell Isolation for Spermatogonial Stem Cell Transplantation in Domestic Cats

INA DOBRINSKI

HANG LEE

MARGARET C. MCENTEE

From the ^* James A. Baker Institute for Animal Health, College of Veterinary Medicine, Cornell University, Ithaca, New York; the Center for Animal Transgenesis and Germ Cell Research, Department of Clinical Studies, New Bolton Center, School of Veterinary Medicine, University of Pennsylvania, Kennett Square, Pennsylvania; the Conservation Genome Resource Bank for Korean Wildlife, College of Veterinary Medicine and School of Agricultural Biotechnology, Seoul National University, Seoul, South Korea; and the Department of Clinical Sciences, College of Veterinary Medicine, Cornell University, Ithaca, New York.

Correspondence to: Alexander J. Travis, The James A. Baker Institute for Animal Health, College of Veterinary Medicine, Cornell University, Ithaca, NY 14853 (e-mail: ajt32{at}cornell.edu).

The loss of genetic diversity poses a serious threat to the conservation of endangered species, including wild felids. We are attempting to develop spermatogonial stem cell transplantation in the cat as a tool to preserve and propagate male germ-plasm from genetically valuable animals, be they threatened wild species or lines of cats used as models for inherited diseases. In this study, we investigated the use of local external beam radiation treatment to deplete the endogenous germ cells of male domestic cats, a step necessary to prepare them for use as recipients for transplantation. Testes of 5-month-old domestic cats were irradiated with a fractionated dose of 3 Gy per fraction for 3 consecutive days. These cats were castrated at 2, 4, 8, 16, and 32 weeks posttreatment, and progress of spermatogenesis was evaluated histologically and compared against age-matched controls. Even at the latest time points, less than 10% of tubules contained germ cells at any stage of meiosis, showing the efficacy of this protocol. In addition, male germ cells were isolated from the testes of domestic cats using a 2-step enzymatic dissociation to establish a protocol for the preparation of donor cells. The presence and viability of spermatogonia within this population were demonstrated by successful transplantation into, and colonization of, mouse seminiferous tubules. The success of these protocols provides a foundation to perform spermatogonial stem cell transplantation in the domestic cat.

     Key words: Testis, male, radiation, feline, conservation

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