Male Pronuclear Formation using Dog Sperm Derived from Ectopic Testicular Xenografts, Testis, and Epididymis

Background: Testis tissue xenografting and the resultant sperm in a xenograft may provide a unique approach to rescue the genetic material of males that die prematurely and is a model for the study of human spermatogenesis and can represent an alternative approach for fertility preservation in cancer patients. This study was aimed to evaluate the xenogenic dog sperm in formation of male pronucleus following injection into the sheep oocytes. Methods: The in vitro matured slaughterhouse derived sheep oocytes were subjected to Intracytoplasmic Sperm Injection (ICSI) with epididymal, testicular, and xenogenic dog sperm. The ICSI was performed after scoring of the sperm midpiece using an IX71-Olympus inverted microscope with Nomarsky optics. Within 1 hr after injection, the injected oocytes in activated group were exposed to 5 µM ionomycin for 5 min. The data were analyzed by Chi-square and ANOVA using SigmaStat, version 3.5, and p<0.05 was considered significant. Results: The formation of female pronucleus after ICSI of xenogenic sperm was higher than epididymal and testicular sperm in non-activated oocytes. The corresponding rate in activated oocytes was higher or comparable with testicular and epididymal sperm. The rate of male pronucleus formation after ICSI of xenogenic sperm was comparable with injection of two other sperm sources. Oocyte activation had an inductive role in female and male pronuclear formation. Conclusion: Dog xenogenic sperm was capable to induce oocyte activation and proportion of male pronucleous formation was comparable to the testicular and epididymal sperm.