Ultrastructure of human ovarian tissues and risk of cancer cells re-implantation after transplantation to chick embryo chorioallantois membrane (CAM) following vitrification or slow freezing

Ovarian follicle depletion and premature ovarian failure are significant challenges in cancer patients subjected to radio- or chemotherapy. Ovarian tissue (OT) cryopreservation would be an option when other fertility preservation methods are not accessible. This study aimed to analyze the structure and ultrastructure of human OTs transplanted onto chick embryo chorioallantois membrane (CAM) after cryopreservation by vitrification or slow freezing. OTs from 10 cancer patients underwent cryopreservation. CAM transplantation was done on fresh and cryopreserved OTs, to assign samples to nine study groups as follows: 1) FI-FIII = fresh, 5- and 10-days post-CAM transplantation groups; 2) VI-VIII = vitrified, 5- and 10-days post-transplantation vitrified groups; 3) SFI-SFIII: slow frozen, 5- and 10-days post-transplantation slow freezing groups. Proliferation ability, folliculogenesis, and structural and ultrastructure were analyzed. The density of primordial follicles did not change after both freezing methods, but reduced after 5 (P ≥ 0.05) and 10 days (P ≤ 0.05) post-CAM transplantation. The follicular grade significantly decreased in all transplanted tissues (P ≤ 0.0). The proliferation marker increased after cryopreservation, but reduced after transplantation (P ≤ 0.05). TEM evaluation showed better follicular ultrastructure in the fresh group, after transplantation. Stromal ultrastructure appeared more preserved after vitrification compared with slow freezing. There was no sign of malignant cell contamination after transplantation. Some follicular TEM abnormalities were found in both methods of freezing, with a better transplantation rate after vitrification. Also, enhanced follicular activation resulted in faster follicular depletion in this method. The information regarding post grafting events would improve our knowledge for longer OTs' lifespans.