Elisabeth Jeremias

Restoration of ovarian function after autotransplantation of intact frozen-thawed sheep ovaries with microvascular anastomosis

OBJECTIVE To test the feasibility of transplanting an intact frozen-thawed ovary with microvascular anastomosis of the ovarian vascular pedicle to the deep inferior epigastric vessels. DESIGN Chronic survival study. SETTING Biological Resources Unit, The Cleveland Clinic Foundation. ANIMAL(S) Adult merino ewes. INTERVENTION(S) Bilateral laparoscopic oophorectomy was performed on 17 synchronized ewes. In one group of animals (Group I, n = 11), both ovaries were cryopreserved intact with their vascular pedicles. In another group of animals (Group II, n = 6), ovarian cortical strips were prepared from each ovary and cryopreserved. After thawing, follicular viability and apoptosis rates were assessed using one ovary. The other ovary was transplanted to the abdominal wall with microvascular anastomosis (Group I). In Group II, the ovarian cortical strips were placed in the anterior abdominal wall. Ovaries were harvested after 8-10 days in situ and subjected to histological evaluation. MAIN OUTCOME MEASURE(S) Blood flow, apoptotic signals, follicular viability, serum estradiol (E(2)), follicle-stimulating hormone (FSH), and histology. RESULT(S) No significant differences were found in the mean values of apoptosis (mostly in the atretic and some secondary follicles) and follicular viability in both groups. In Group I, immediate and long-term patency were documented in 100% and 27% (3/11) of the grafts, respectively; and postoperative FSH levels were similar to preoperative values in animals with patent vessels. In Group II, postoperative FSH levels were significantly higher than the preoperative ones (P=.03). CONCLUSION(S) Transplantation of an intact frozen-thawed ovary is technically feasible. Using this approach, immediate restoration of vascular supply and ovarian hormonal functions is possible.

Heterotopic autotransplantation of the ovary with microvascular anastomosis: A novel surgical technique

OBJECTIVE To test the feasibility of transplanting an entire ovary with anastomosis of the ovarian vascular pedicle. DESIGN Long-term survival study. SETTING Biological Resources Unit, Cleveland Clinic Foundation. ANIMAL(S) Five adult, nonpregnant ewes. INTERVENTION(S) Laparoscopic bilateral oophorectomy was performed. Ovaries were autotransplanted into the abdominal wall, and microsurgical vascular anastomosis of the ovarian to the inferior epigastric vessels was performed. The transplant was removed and evaluated after 7 +/- 1 days. MAIN OUTCOME MEASURE(S) Blood flow, serum E2 and FSH levels, and histologic characteristics. RESULT(S) At follow-up three transplants were viable; they showed no signs of necrosis, and patency of the vascular anastomosis was confirmed. Serum E2 levels did not change significantly after transplantation in the patent vessel group (155.3 +/- 46.1 vs. 125.7 +/- 44.6 pg/mL) or the nonpatent vessel group (99 vs. 158 pg/mL). Serum FSH level in the patent vessel group did not change significantly from before to after transplantation (70.6 +/- 37.2 ng/mL vs. 95.1 +/- 17.7 ng/mL), whereas a large increase in FSH level was observed in the nonpatent vessel group (52.3 ng/mL vs. 522 ng/mL). The patent vessel group had significantly more follicles after transplantation than did the nonpatent vessel group (6 +/- 1 vs. 1 +/- 1). CONCLUSION(S) In conjunction with improved protocols for cryopreservation, ovarian autotransplantation with vascular anastomosis may be superior to current ovarian tissue banking and grafting techniques.

Assessment of tissue injury in cryopreserved ovarian tissue

Ovarian tissue banking is a developing technique aimed to preserve fertility in women at risk for premature ovarian failure (1–3). It entails cryopreservation of ovarian tissue with the option of autografting. Reports on the autografting of cryopreserved ovarian cortex segments describe the resumption of steroid production (4), follicle development (5, 6), and (in sheep) model successful pregnancies (7, 8). However, tissue loss due to procedural steps is expected to have detrimental effects on the reproductive potential of frozen-thawed ovarian grafts. We conducted a study in a porcine model to investigate how freezing and thawing affects ovarian tissue and whether warm ischemia time before the cryopreservation process has any added detrimental effects.

Histological evaluation and in situ localization of apoptosis in fresh and cryopreserved ovarian tissue

Objective: To study the feasibility of using combined morphology and terminal deoxynucleotidyl transferase-mediated dUTP-digoxigenin nick-end labeling (TUNEL) for apoptosis detection and the impact of cryopreservation on this process. Materials and methods: We conducted this investigation using a porcine animal model. Bilateral oophorectomy was performed in eight sows. The ovarian tissues were divided into two parts; one part was immediately fixed while the other was cryopreserved. The cryopreserved specimens were subsequently thawed and then fixed. All the specimens were sectioned, fixed and stained with HE 2) the nuclei of the granulosa cells, but not those of theca or stromal cells, were TUNEL positive; 3) some cells with histological features of necrosis and apoptosis were TUNEL negative, and 6) The distribution of apoptosis was not different between cryopreserved tissue and freshly fixed tissue. Conclusions: the presence of apoptosis in the atretic follicles may suggest its involvement in follicular atresia; and 2) combined histology and TUNEL assay may be a useful method for detection of apoptosis.

Autologous transplantation of cryopreserved ovary induces the generation of antiovary antibodies in sheep.

Ovarian cryopreservation and grafting has resulted in successful ovulation, pregnancy, and live-born animals including mice and sheep (1, 2). The procedure is currently being offered to humans as a new method of assisted reproduction. However, its value frequently has been questioned because of a potential to reintroduce the malignant cells, viral infections, and induction of immune responses against the transplanted ovarian tissue (3, 4).

Does autologous transplantation of cryopreserved ovary result in induction of anti-ovarian antibodies?

OVARY RESULT IN INDUCTION OF ANTI-OVARIAN ANTIBODIES? Navid Esfandiari, Mohamed A Bedaiwy, Ashok Agarwal, Elisabeth Jeremias, Rakesh K Sharma, Tommaso Falcone, Cleveland Clinic Foundation Objective: Cryopreservation and autologous transplantation of ovary has been recognized as an encouraging technique for preservation of reproductive function and fertility for female cancer patients. Ovarian tissue in human and some laboratory animals has been cryopreserved and autografted for some time, but there are no studies on the immunological consequences following ovarian transplantation. In this study using sheep as a model, we examined whether orthotopic transplantation of cryopreserved entire ovary or ovarian cortical strips can result in antibody formation against ovarian tissue antigens.