Abstract
BackgroundOvarian tissue vitrification is an alternative fertility preservation procedure for young female patients prior to gonadotoxic treatment. Primordial follicle loss might be a potential issue for vitrification and transplantation procedures. This study aimed to evaluate primordial follicle density and deoxyribonucleic acid (DNA) fragmentation in each stage of the preservation procedure of goat ovarian tissue. Follicle density and DNA fragmentation were examined microscopically after staining with hematoxylin eosin and TUNEL assay, respectively. Both parameters were compared between fresh, fresh-transplanted, vitrification, and vitrification-transplanted groups.ResultsA significant decrease was observed in the primordial follicle proportion after vitrification and transplantation compared to the primordial follicle proportion in the fresh group (88.09% vs 52.42%, p < 0.05, 95% CI 11.54, 66.94). There was no significant difference in DNA fragmentations of primordial follicles between each group (p > 0.05).ConclusionsThe vitrification and transplantation process of goat ovarian strips could cause the primordial follicles loss and DNA damage of the follicles. However, primordial follicles loss and DNA damage were not significantly different in each procedure.
Highlights
Ovarian tissue vitrification is an alternative fertility preservation procedure for young female patients prior to gonadotoxic treatment
Ovarian tissue vitrification-thawing and transplantation We developed the vitrification method based on modifications of the method described by Suzuki et al in 2015 [19]
Histological analysis Follicle density In order to determine the effects of vitrification and transplantation of ovarian strips on the development of primordial follicles and deoxyribonucleic acid (DNA) fragmentation, primordial follicles were counted after 5 days of incubation of the ovarian strips on chorioallanthoic membrane (CAM) and analyzed histologically after hematoxylin and eosin (HE) staining (Fig. 2)
Summary
Ovarian tissue vitrification is an alternative fertility preservation procedure for young female patients prior to gonadotoxic treatment. This study aimed to evaluate primordial follicle density and deoxyribonucleic acid (DNA) fragmentation in each stage of the preservation procedure of goat ovarian tissue. Follicle density and DNA fragmentation were examined microscopically after staining with hematoxylin eosin and TUNEL assay, respectively. Both parameters were compared between fresh, fresh-transplanted, vitrification, and vitrification-transplanted groups. Loss of primordial follicles can occur during the preparation, cryopreservation, and retransplantation steps. Primordial follicle loss occurs due to fragmentation of ovarian tissue which could disrupt Hippo signaling in granulosa cells through activation PI3K/Akt/mTORC1 pathway in oocytes [9–11].
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