Function of Cryopreserved Cat Ovarian Tissue after Autotransplantation

SIMPLE SUMMARY: Assisted reproduction techniques are potentially important tools for the creation of gene banks largely focused on preserving female germ cells and tissues, cryopreservation being one of the most important. Since there is not yet a protocol established for the preservation of cat ovarian tissue, we decided to assess our cryopreservation protocol with autotransplantation of the ovary. Our study showed that even though follicular survival was low, follicles were able to survive up to 28 days of transplantation and develop up to the antral stage, which helps elucidate the path for preservation of felid ovaries. Once this technique is improved, it may contribute to the preservation of wild feline species. ABSTRACT: The aim of this study was to assess a slow-freezing protocol of cat ovarian tissue cryopreservation using autotransplantation. Four adult queens were ovariohysterectomized and the ovaries were fragmented and cryopreserved. After one week, the grafts were thawed and autografted to the subcutaneous tissue of the dorsal neck of each queen, then randomly removed after 7, 14, 28, 49, and 63 days after transplantation. Percentages of morphologically normal primordial and growing follicles (MNFs) were 88% and 97%, respectively, in fresh tissue samples (fresh controls), and 74% and 100%, respectively, immediately after thawing (cryo D0). No MNFs were found after 49 days of transplantation. In both fresh control and cryo D0 fragments, granulosa cells were frequently in proliferation. Two morphologically normal antral follicles were detected in one queen on Day 28 post-transplantation. Connective tissue fibers increased, suggesting replacement of active ovarian cortex by fibrous tissue. Tissue vascularization was observed at 7 days after grafting, and wide blood vessels were clearly visible on Days 49 and 63. In conclusion, although follicular survival was low after cryopreservation and grafting of cat ovarian tissue, follicles were able to develop up to the antral stage, which is an encouraging outcome.