Ovarian function 6 years after cryopreservation and transplantation of whole sheep ovaries.

Whole ovary cryopreservation and transplantation has been proposed as a method for preserving long-term ovarian function. This work reports ovarian function 6years post transplantation of frozen-thawed whole sheep ovaries. Three 9-month-old Assaf sheep underwent unilateral oophorectomy to provide organs for the experiments. After perfusing with cold University of Wisconsin solution supplemented with 10% dimethyl sulphoxide, ovaries were cryopreserved using unidirectional solidification freezing technology. After thawing, ovaries were re-perfused and re-transplanted orthotopically by microvascular re-anastomosis, to the contralateral ovarian pedicle after removing the remaining ovary. Six years following transplantation and after inducing superovulation, the sheep were killed and the ovaries analysed. Two ovaries had normal size and shape showing some recent corpora lutea, while the third showed atrophic changes. A total of 36 antral follicles were counted by transillumination and four germinal vesicle oocytes were aspirated and matured in vitro to metaphase II. Serum progesterone concentrations were indicative of ovulatory activity in one of the three sheep. Histological evaluations revealed normal tissue architecture, intact blood vessels and follicles at various stages. Currently, this is the longest recorded ovarian function after cryopreservation and re-transplantation. Cryopreservation of whole ovaries, using directional freezing combined with microvascular anastomosis, is a promising method for preserving long-term reproductive capacity and endocrine function.

Oocyte recovery, embryo development and ovarian function after cryopreservation and transplantation of whole sheep ovary.

BACKGROUND: Successful cryopreservation of a whole ovary may provide a solution for women with premature ovarian failure. The aim of this study was to evaluate the function of cryopreserved whole sheep ovaries both in vitro and in vivo. METHODS: Transplantation of frozen-thawed intact ovaries was performed on eight sheep by artery and vein anastomosis to the contralateral ovarian artery and vein. The remaining ovary was removed. Oocyte aspiration was performed 1 and 4 months post-transplantation. Serum progesterone levels were measured after 24 and 36 months. Magnetic resonance imaging (MRI) was carried out 12 months after transplantation. RESULTS: Progesterone activity was detected in three sheep from 24 to 36 months post-transplantation. Oocyte retrieval was successful in two sheep and parthenogenic activation has resulted in embryonic development up to the 8-cell stage. MRI revealed an intact ovary with small follicles and intact blood vessels. CONCLUSIONS: Whole ovaries, and the follicles and blood vessels they contain, are able to survive cryopreservation. In addition, MRI has shown that blood vessels were intact and that normal blood flow had resumed to the transplant. We conclude that immediate and long-term hormonal restoration and normal ovulation is possible after cryopreservation and transplantation of whole ovaries in sheep.