[01] Mohammad Sadra Shirazi, Reproductive Biotechnology Research Center, Avicenna Research Institute (ACECR), Tehran, Iran. [02] Banafsheh Heidari, Reproductive Biotechnology Research Center, Avicenna Research Institute (ACECR), Tehran, Iran. [03] Mohammad Mehdi Naderi, Reproductive Biotechnology Research Center, Avicenna Research Institute (ACECR), Tehran, Iran. [04] Bahareh Behzadi, Reproductive Biotechnology Research Center, Avicenna Research Institute (ACECR), Tehran, Iran. [05] Ali Sarvari, Reproductive Biotechnology Research Center, Avicenna Research Institute (ACECR), Tehran, Iran. [06] Sara Borjian-Boroujeni, Reproductive Biotechnology Research Center, Avicenna Research Institute (ACECR), Tehran, Iran. [07] Morteza Farab, Reproductive Biotechnology Research Center, Avicenna Research Institute (ACECR), Tehran, Iran. [08] Abolfazl Shirazi, Reproductive Biotechnology Research Center, Avicenna Research Institute (ACECR), Tehran, Iran;Research Institute of Animal Embryo Technology, Shahrekord University, Shahrekord, Iran.

Assisted reproductive techniques involving isolation, culture, and transplantation of spermatogonial stem cells offer unique approach to manipulate the male germline. The application of these techniques in farm animals has been the subject of an increasing number of studies, mostly because of its potential as an alternative strategy in producing transgenic livestock with higher efficiency and less time and capital requirement than the current methods. The aim of this study was to assess the colonization and differentiation potentials of enriched goat spermatogonia into the mouse testes. Moreover, because stem cells may need to be preserved for several years before re-introduction to the recipient testes, we developed the efficient cryopreservation technique for type A spermatogonoa. The enzymatically isolated SSCs obtained from one month old goats’ testes were enriched by using discontinuous percoll density, and followed by cryopreservation protocol. After xenotransplantation of prepared goat testicular cells into the mouse rete testis, the proliferative activity and stemness potential of SSCs were evaluated and compared with in vitro culture condition. We demonstrated that the viability of testicular cells after cryopreservation was significantly lower than fresh cells, although these cells had normal structural and functional characteristics (P <0.001). Donor goat spermatogonia were able to survive and colonize in depleted recipient’s testis at 80 days after transplantation, but later stages of donor-derived spermatogenesis were not observed at this time. Although cross-species spermatogonial transplantation did not have the envisioned immediate practical application, it nonetheless provides a bioassay for stem cell potential of germ cells isolated from other species.

Cryopreservation, Goat, Mouse, PGP9.5, Spermatogonial Stem Cells, Transplantation

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