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Hum. Reprod. Advance Access originally published online on December 13, 2006
Human Reproduction 2007 22(4):1060-1067; doi:10.1093/humrep/del471
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© The Author 2006. Published by Oxford University Press on behalf of the European Society of Human Reproduction and Embryology. All rights reserved. For Permissions, please email: journals.permissions@oxfordjournals.org

Effect of cold storage and cryopreservation of immature non-human primate testicular tissue on spermatogonial stem cell potential in xenografts

Kirsi Jahnukainen1,2,3,4, Jens Ehmcke1, Scott D. Hergenrother1 and Stefan Schlatt1

1 Department of Cell Biology and Physiology, Center for Research in Reproductive Physiology, University of Pittsburgh, School of Medicine, Pittsburgh, PA, USA 2 Pediatric Endocrinology Unit, Department of Woman and Child Health, Karolinska Institute and University Hospital, Stockholm, Sweden 3 Department of Pediatrics, University of Turku, Turku, Finland

4 To whom correspondence should be addressed at: Department of Pediatrics, University of Turku, FIN-20520 Turku, Finland. E-mail: kirsi.jahnukainen{at}ki.se

BACKGROUND: Successful cryopreservation of gonadal tissue is an important factor in guaranteeing the fertility preservation via germ cell or testicular tissue transplantation. The aim of this study was to evaluate the effects of cooling and cryopreservation on spermatogonial stem cell survival and function of immature non-human primate testicular tissue xenografted to nude mice.

METHODS: Group 1 (control group) received subcutaneous grafts of fresh immature rhesus monkey testes. The treatment groups received grafts after 24 h cooling in ice-cold medium (Group 2), after 24 h of cryopreservation without cryoprotectant (Group 3), with ethylene glycol (Group 4: 1.4 M) or with dimethylsulphoxide (DMSO) (group 5: 1.4 M; group 6: 0.7 M), using cooling rates of 0.5°C/min. The graft number, weight and histology were examined 3–5 months later.

RESULTS: After xenografting, grafts from fresh and cooled tissue showed good survival and spermatogenic induction to spermatocytes. Cryopreservation in 1.4 M DMSO also allowed grafts to initiate spermatogenesis. In contrast, 0.7 M DMSO and ethylene glycol showed inferior protection.

CONCLUSIONS: Our observations suggest that cryopreservation of immature primate testis is a feasible approach to maintain spermatogonial stem cells and may serve as a promising tool for fertility preservation of prepubertal boys. The possibility to delay the transplantation of cooled samples suggests an option for clinical centralization of testicular tissue cryopreservation.

Key words: cryopreservation/prepuberty/primate/testis/xenograft

Submitted on October 4, 2006; resubmitted on October 28, 2006; accepted on November 15, 2006.


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