Autologous Ovarian Transplantation Research Articles

Comparison of orthotopic and heterotopic autologous ovarian tissue transplantation outcomes

To compare the outcomes of orthotopic and heterotopic ovarian tissue transplantation (OTT) techniques. Mixed prospective-retrospective cohort study. Academic hospital. A total of 14 recipients of autologous OTT. Of the 14 women, 12 who received orthotopic (n = 6) or heterotopic (n = 6) transplants met the inclusion criteria. All orthotopic transplants and one heterotopic ovarian tissue transplant were performed laparoscopically. Although 5 of the 6 remaining heterotopic transplants were performed subcutaneously under local anesthesia or intravenous sedation, one was performed with robotic assistance. With the exception of one recipient who solely desired restoration of endocrine function, all underwent oocyte retrieval either to cryopreserve oocytes and embryos before the graft function ceased or because they could not otherwise conceive (hysterectomy, radiation damage, and heterotopic transplant). Primary outcome measures were graft function and longevity, and the number of embryos generated per retrieval. The mean age at ovarian tissue harvesting and transplantation was lower in patients with orthotopic vs. heterotopic transplants, although the proportion of transplanted ovarian cortex was lower in heterotopic transplant cases. All grafts restored ovarian endocrine function. Fertilization rates, the number of embryos generated per retrieval, and the mean number of nonarrested embryos were significantly lower in heterotopic OTT. However, time to function and graft longevity were similar between the groups. Although 4 of the 6 women conceived and delivered 7 children among orthotopic ovarian tissue recipients, one recipient had 3 spontaneous live births after heterotopic OTT, presumably because of the induction of function in the remaining menopausal ovary. It appears that orthotopic OTT results in higher gamete and embryo quality. However, the endocrine function restoration rate and longevity are similar between the 2 approaches. When feasible, orthotopic OTT should be preferred for those who intend to conceive, although a less invasive heterotopic OTT can be performed for those who primarily desire ovarian endocrine function.

Safety of ovarian cryopreservation and transplantation in patients with acute leukemia: a case series

With increased success, ovarian tissue cryopreservation has recently become a standard technique for fertility preservation. However, malignant cell introduction through ovarian tissue transplantation remains a major concern for patients with acute leukemias. This study aimed to investigate the safety of performing autologous ovarian tissue transplantation in survivors of acute leukemia. Clinical, histopathological, and molecular data of 4 women with acute myeloid leukemia and 2 women with acute lymphoblastic leukemia who underwent ovarian tissue cryopreservation and transplantation were analyzed in this case series. Following cryopreservation of 66% to 100% of an ovarian cortex with a slow freezing method, all women received high-dose multiagent alkylating preconditioning chemotherapy for allogeneic hematopoietic stem cell transplantation. Before the ovarian tissue transplantation, (1) antral follicle counts, serum antimüllerian hormone and follicle-stimulating hormone levels were assessed to confirm primary ovarian insufficiency; (2) all recipients were cleared by their hematologist-oncologists; (3) representative cortical strips were screened for leukemia infiltration by histologic (hematoxylin and eosin staining), immunohistochemical (CD3, CD20, CD34, CD68, CD117, CD163, PAX-5, Tdt, lysozyme, and MPO), and molecular marker evaluation (BCR/ABL p190 and AML1/ETO) where appropriate. The median age was 20 years (interquartile range, 15-32) at ovarian tissue cryopreservation. Before undergoing hematopoietic stem cell transplantation, all patients received induction or consolidation chemotherapy that included cytarabine+ daunorubicin or Berlin-Frankfurt-Munich-95 protocol and were in remission. The mean serum antimüllerian hormone was 1.9±1.7 ng/mL before ovarian tissue cryopreservation. In all cases, ovarian tissue screening for leukemic cells was negative. Ovarian transplantation was performed laparoscopically with or without robotic assistance, after a median of 74.5 months (interquartile range, 41-120) after ovarian tissue cryopreservation. Ovarian function resumed in all patients after a median of 3.0 months (range, 2.5-4.0), and 2 women had 1 live birth each. The median graft longevity was 35.5 months (interquartile range, 18-57) after ovarian tissue transplantation. After a median follow-up of 51 months (interquartile range, 20-74), all patients remained relapse-free. In 1 patient, the graft was removed during cesarean delivery and was negative for immunochemical leukemia markers. Our long-term follow-up demonstrated no evidence of disease relapse after ovarian tissue transplantation in patients with acute leukemia who received allogeneic hematopoietic stem cell transplantation. This safety profile may be explained by the fact that these patients are induced into remission by nongonadotoxic induction chemotherapy before undergoing ovarian tissue cryopreservation. We propose that ovarian tissue cryopreservation should not be excluded as a fertility preservation option for young women with leukemia who are due to receive preconditioning chemotherapy before allogeneic hematopoietic stem cell transplantation.

IS IT SAFE TO PERFORM OVARIAN CRYOPRESERVATION AND TRANSPLANTATION IN PATIENTS WITH LEUKEMIA A 5-YEAR SURVIVAL ANALYSIS DATA

Although ovarian tissue cryopreservation is performed in patients with leukemia undergoing Allogenic Hematopoietic Stem Cell Transplantation (Allo-HSCT), auto-transplantation of these tissues have raised concerns with the risk of reintroducing cancer cells. We performed this retrospective study, spanning 14 years to investigate the safety of performing autologous ovarian tissue transplantation in acute leukemia survivors.

Updating the recommendations of the Working Group for the Preservation of Fertility in Oncological and Hematological Patients and Other Patients Treating Gonadier Therapies "ONCOFERTILITY" (GROF) of the Polish Society of Oncological Gynecology regarding cryopreserves and autologous transplant.

Update of the recommendations of the Fertility Preservation Working Group in Oncological, Hematological and Other Patients Treated with Gonadotoxic Therapies "ONCOFERTILITY" (GROF) of the Polish Society of Oncological Gynecology regarding cryopreservation and autologous ovarian tissue transplantation. The Fertility Preservation Working Group in Oncological, Hematological and Other Patients Treated with Gonadotoxic Therapies "ONCOFERTILITY" (GROF) of the Polish Society of Oncological Gynecology has developed current clinical guidelines and recommendations to improve the quality of healthcare provision in the area of reproductive health in patients undergoing therapy that may impair their reproductive potential. The guidelines are based on current scientific evidence available at the time of writing this document. In the absence of scientific evidence on some aspects, a consensus was reached among GROF stakeholders. The purpose of the guidelines is to assist healthcare professionals in making decisions in specific clinical situations regarding the selection of an appropriate and effective diagnostic and therapeutic process. The document provides practical guidelines for the management of cryopreservation and autologous ovarian tissue transplantation.

Effect of resveratrol on mouse ovarian vitrification and transplantation

BackgroundAfter ovarian tissue transplantation, ischemia-reperfusion injury and free radicals cause follicle depletion and apoptosis. Therefore, the use of antioxidants to reduce the production of free radicals is an important method to address the consequences of ischemia-reperfusion injury. Resveratrol is a natural active polyphenol compound with anti-inflammatory, antitumor, strong antioxidant and anti-free radical properties. The aim of this study was to investigate whether resveratrol could improve the effect of autologous ovarian transplantation after cryopreserve-thawn mouse ovarian tissue.MethodsWhole-ovary vitrification and autotransplantation models were used to investigate the effects of resveratrol. Six-week-old female mice from the Institute of Cancer Research (ICR) were subjected to vitrification. All ovaries were preserved in liquid nitrogen for 1 week before being thawed. After thawing, ovarian tissues were autotransplanted in the bilateral kidney capsules. Mice (n = 72) were randomly divided into four groups to determine the optimal concentration of resveratrol (experiment I).Treatments were given as follows: saline, 5 mg/kg resveratrol, 15 mg/kg resveratrol and 45 mg/kg resveratrol, which were administered orally for one week. Grafted ovaries were collected for analysis on days 3, 7, and 21 after transplantation. Ovarian follicle morphology was assessed by hematoxylin and eosin staining. Serum FSH and E2 levels were measured to estimate the transplanted ovarian reserve and endocrine function. Other mice were randomly divided into two groups—saline and 45 mg/kg resveratrol to further evaluate the effect of resveratrol and explore the mechanisms underlying this effect (experiment II). Ovarian follicle apoptosis was assessed by terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL) assays. Immunohistochemistry, qRT-PCR and western blotting (MDA, SOD, NF-κB, IL-6 and SIRT1) were used to explore the mechanisms of resveratrol. Moreover, oocytes derived from autotransplanted ovaries at 21 days were cultured and fertilized in vitro.ResultsThe proportions of morphologically normal (G1) follicles at 3, 7 and 21 days were significantly higher in the 45 mg/kg resveratrol group than in the saline group. The TUNEL-stained follicles (%) at 7 days were significantly decreased in the 45 mg/kg resveratrol group compared with the saline group. Western blot analysis revealed that SOD2 and SIRT1 levels were significantly higher in the 45 mg/kg resveratrol group than in the saline group at day 7 and that MDA and NF-κB levels were lower in the saline group on day 3. Likewise, IL-6 was lower in the saline group on day 7. These results are basically consistent with the qRT-PCR results. In addition, the mean number of retrieved oocytes and fertilization and cleavage were significantly increased in the 45 mg/kg resveratrol group compared with the saline group.ConclusionsAdministration of resveratrol could improve the quality of cryopreserved mouse ovarian tissue after transplantation and the embryo outcome, through anti-inflammatory and antioxidative mechanisms.

Sexuality- and Fertility-Related Issues in Women after Allogeneic Hematopoietic Stem Cell Transplantation

Sexual dysfunction and fertility related issues appear as major post-allogeneic hematopoietic stem cell transplantation (HSCT) late effects in young women, with a heavy impact on quality of life. The objective of the present study was to evaluate the impact of disease and treatments on sexual quality of life, ovarian function, and family planning initiatives in the context of allogeneic HSCT. Between January 2014 and January 2016, adult female patientswho underwent HSCT before age 35 and had been followed formore than 2 years in our centerwere offered participation inthe study through a self-reported surveyand/or ovarian function assessment if age <40 at inclusion. A total of 63 patients were included, with a median age of 23.4 years at transplantation and 30.9 years at inclusion. Twenty-nine patients (46%) underwent HSCT for acute leukemia and 16 (25%) underwent HSCT for aplastic anemia (AA). The conditioning regimen was myeloablative conditioning (MAC) in 37 patients (59%) and reduced-intensity conditioning (RIC) in 26 (41%). Fifty-eight patients completed the survey, and 34 were evaluated for ovarian function. Symptoms of hypoestrogenism were reported by 86% of the patients and changes in sexual life were reported by 76%, due mainly to low sex drive, negative impact of infertility problems, physical sequelae, and loss of self-confidence. Premature ovarian failure (POF) occurred in 74% of patients and was significantly associated with conditioning regimen (MACversusRIC; P=.001) and baseline disease (bone marrow failureversusacute leukemiaversusothers; P < .001). However, one-half of the patients developed a POF despite the use of a RIC regimen. For 27 patients (47%), disease and treatments modified their desire for pregnancy, due mainly to fear of relapse and of disease transmission to offspring. Thirteen pregnancies were reported (21%), of which 8 were spontaneous and 5 were obtained through assisted reproductive technologies, mainly oocyte donation. With a median post-transplantation follow-up of 12.2 years, the 10-year cumulative incidence of first pregnancy was 16.6% (95% CI, 8.8-30.0). Among 20 patients (32%) who engaged in a family planning initiative, 13 (65%) succeeded in having children: 11 got pregnant and 2 adopted. Sixteen patients benefited from fertility preservation techniques consisting of ovarian tissue cryopreservation, and a single autologous ovarian tissue transplantation had been performed at the time of this report. This study shows a strong impact of disease and treatments on sexual quality of life, ovarian function, and family planning initiatives in the context of HSCT. It demonstrates the need to improve clinicians' awareness of sexual health- and fertility-related issues after HSCT. The difficulty of predicting ovarian function and fertility issues after RIC supports wide indications of pretransplantation fertility preservation. Evaluation of the use of cryopreserved ovarian tissues is warranted.

Advances in preservation of endocrine function with ovarian tissue cryopreservation and transplantation in cervical cancer.

The incidence of cervical cancer ranks the first among the gynecological malignant tumors. Given the implementation of widespread cervical cancer screening and the improvement of clinical treatment, the cervical cancer mortality has decreased substantially. However, surgery, radiotherapy, and chemotherapy may impair the ovarian function, resulting in premature ovarian insufficiency and premature climacteric, which has reduced the quality of life in young patients. As a new fertility preservation technology, autologous ovarian tissue cryopreservation and transplantation is developing rapidly and showing great potentiality in preserving ovarian endocrine function of young cervical cancer patients.

A Novel Treatment of Symptomatic Menopause Autologous Ovarian Transplantation Research in Progress

A Novel Treatment of Symptomatic Menopause Autologous Ovarian Transplantation Research in Progress

Robot-assisted orthotopic and heterotopic ovarian tissue transplantation techniques: surgical advances since our first success in 2000

To demonstrate the technical advances since the time we reported the first successful case in 2000 and our modern approach to autologous transplantation of frozen-thawed human ovarian tissue. A step-by-step video demonstration of three surgical approaches was created by editing the surgical footage obtained during ovarian transplantation procedures. Academic. Three patients who previously underwent ovarian tissue harvesting and cryopreservation before gonadotoxic cancer treatments or radical cancer surgery are presented. The illustrated techniques include robot-assisted orthotopic (technique 1) and heterotopic (technique 2) approaches using the da Vinci Xi (Intuitive Surgical) robotic system and a decellularized human extracellular tissue matrix (Alloderm; LifeCell Corp.) as a tissue scaffold, as well as a percutaneous autotransplantation approach (technique 3). Successful completion of procedures without complications and ovarian graft function with demonstration of E2 production and follicle development. All cases were completed without complications. Ovarian graft function was confirmed by E2 production, follicle growth by 10-14weeks after transplantation, and later embryo development. Since our first report of successful restoration of ovarian function after orthotopic transplantation of frozen-banked ovarian tissue in 2000 (1), followed by our first reports of subcutaneous heterotopic transplantation techniques (2, 3), ovarian tissue cryopreservation followed by subsequent transplantation has become a promising fertility preservation option for young women with cancer who do not have sufficient time to undergo oocyte or embryo cryopreservation and for prepubertal girls (4, 5). The same approach also has the advantage of restoring ovarian endocrine function and fertility without a need for assisted reproduction (6, 7). In the very first successful procedure that we reported in 2000, we used conventional laparoscopy, and the tissues were reconstructed and mounted on a polycellulose scaffold (Surgicel) (1, 7). Since then, we have made significant modifications in our surgical approach with potential improvements in outcomes. Here we illustrate three main techniques of ovarian tissue transplantation resulting in the restoration of ovarian function in all cases. In the first two cases, we illustrated the robot-assisted orthotopic and heterotopic approaches using Alloderm. Robotic ovarian transplantation may increase precision, provide more delicate graft handling, and reduce the time from tissue thawing to transplantation (6, 8). Alloderm is regenerated de-epithelized human cadaver skin, which consists of several extracellular matrix components. It has been safely used in the surgery and dentistry fields for enhancing tissue regeneration and vascularization (9, 10). Furthermore, our earlier laboratory work indicated the critical role of extracellular matrix in primordial follicle growth initiation and preantral follicle growth (11, 12). Prior to our use of Alloderm as part of ovarian transplant procedures, we tested it in human ovarian xenograft models and found Alloderm to incorporate well with ovarian tissue (8). Only after that test did we adopt it for use in ovarian transplants. The utility of the extracellular tissue matrix may thus enhance our ovarian autotransplantation techniques by facilitating ovarian reconstruction and potentially improving neovascularization. In fact, we have seen improved follicle growth and response to ovarian stimulation with the use of Alloderm in our first cases (8). We use heterotopic ovarian transplantation when the pelvis is not suitable for autotransplantation due to past radiation or scarring or when there are other medical contraindications for transplantation in the pelvis. The third technique we illustrated was percutaneous heterotopic ovarian autotransplantation. This is a simple approach that can be used in surgically high-risk patients, as it is done with local anesthesia or IV sedation and without entering abdominal cavity. Additionally, same approach can be utilized when there is heightened concern that the ovarian tissue may harbor a disease that can recur, requiring close surveillance and easier removal of the ovarian graft. While ovarian endocrine function and follicle growth are restored with efficiency using the percutaneous ovarian transplants, our initial experience suggests that oocyte quality may be impaired in SC locations (2, 3, 13). Hence that technique may be more suitable when the only purpose is restoration of ovarian endocrine function. However, we have encountered recurrent live births from spontaneous conceptions following SC ovarian transplants, prompting the question of whether the grafted tissue can augment the function of in situ menopausal ovary (13, 14). While ovarian cryopreservation and transplantation may no longer be considered experimental, there are many exciting questions remaining to be answered on the full potential of this procedure.

Current Success and Efficiency of Autologous Ovarian Transplantation: A Meta-Analysis.

Ovarian cryopreservation followed by autotransplantation is still considered an experimental strategy for fertility preservation (FP) mainly because the success rates are unknown. To determine cohort epidemiologic characteristics and success rates of autologous ovarian tissue transplantation (OTT) with previously cryopreserved tissue. Literature review from 1999 to October 1, 2016. Additional cases were retrieved from meeting abstracts and own database. We selected studies that reported autologous OTT with previously banked tissue in humans. We did not include any cases involving fresh ovarian tissue transplantation or those performed to treat idiopathic premature ovarian failure/insufficiency. Both authors reviewed and selected studies for eligibility, which resulted in 59 full-text studies assessed for eligibility. Cases were extracted from original reports and reviews by the junior author, and the senior author reviewed and verified the extracted data. Nineteen reports were included for qualitative synthesis. In 10 studies, detailed data were available to determine clinical and live birth + ongoing (LB + OG) pregnancy as well as endocrine restoration rates. Three hundred nine OTTs were performed with cryopreserved tissue, resulting in the birth of 84 children and 8 OG pregnancies. The cumulative clinical and LB + OG rates were 57.5% and 37.7%, respectively, and the endocrine restoration rate was 63.9%. Success rates with cryopreserved OTT have reached promising levels. Given these recent data, ovarian tissue cryopreservation should be considered as a viable option for FP.

Robot-assisted Laparoscopic Transplantation of Frozen-thawed Ovarian Tissue

Study ObjectiveTo show a new approach for orthotopic human ovarian tissue transplantation via robot-assisted laparoscopic surgery. DesignA step-by-step video explanation of the surgical technique (Canadian Task Force classification III). SettingAcademic medical center. InterventionsThe robot-assisted transplantation approach consisted of 3 steps: (1) reconstruction of the ovarian tissue graft, (2) preparation of the contralateral menopausal ovary as the recipient site, and 3) transplantation of the reconstructed graft to the bivalved contralateral ovary. Institutional review board approval was obtained. Measurements and Main ResultsAlthough still experimental, cryopreservation and subsequent transplantation of frozen-thawed ovarian tissue are currently the only available methods for prepubertal girls and young women with cancer who are not eligible for established fertility preservation options such as oocyte or embryo cryopreservation [1]. We performed the first reported autologous ovarian transplantation with a conventional laparoscopic technique [2]. To date, over 60 babies have been born after the orthotopic transplantation of cryopreserved ovarian tissue, and this number is growing [3,4]. Until recently, all of these children were born from ovarian transplants that were performed via laparotomy or conventional laparoscopy [5]. We have recently developed a robot-assisted ovarian transplantation procedure that uses an extracellular matrix scaffold to facilitate ovarian reconstruction, handling, and revascularization. Both of the procedures resulted in robust ovarian function and births [6]. The purpose of this video reports the surgical technique in detail, which uses the da Vinci Xi (Intuitive Surgical Inc, Sunnyvale, CA) robotic system for transplantation, and a decellularized human extracellular tissue matrix (Alloderm; LifeCell Corp, Branchburg, NJ) for graft reconstruction. ConclusionRobotic ovarian transplantation may have several advantages, which include precision, more delicate graft handling, and reduced time from tissue thawing to transplantation. The collective usefulness of the extracellular tissue matrix may enhance this technique by enabling a niche for ovarian reconstruction and potentially enhanced revascularization. The feasibility and comparative advantages of this technique are currently being studied in ongoing trials.

Current succes and efficiency of autologous ovarian transplantation with cryopreserved tissue: a meta-analysis

Ovarian cryopreservation is still considered an experimental fertility preservation strategy and the success rates with the transplantation (OT) of previously banked ovarian tissue are unknown. Here we sought to meta-analyze published and unpublished data, including our own to determine success rates of OT.

Laparoscopic autologous and heterologous ovarian transplantation in orthotopic and heterotopic location without vascular anastomosis in rabbits: morphologic and endocrinologic assessment

The purpose of this study was to assess the viability of the ovarian function and compare the success rate of autologous and heterologous ovarian transplantation in orthotopic and heterotopic location in rabbits. Eighty-nine virgin adult white New Zealand female rabbits weighing 3350–4550 g and 4–5 months old were used. The animals were randomly allocated into five groups: group Α (control group) with bilateral ovariectomy (n = 10); group Β, which underwent ovariectomy and intraperitoneal autologous heterotopic transplantation of the ovaries without vascular pedicle (n = 22); group C, in which ovariectomy was performed and autologous orthotopic transplantation in the same location without vascular anastomosis was performed, (n = 20); group D, included animals with ovariectomy and intraperitoneal heterologous heterotopic transplantation of the ovaries without vascular pedicle (n = 19); and group E, included rabbits which underwent ovariectomy and heterologous orthotopic transplantation in the same location without vascular anastomosis (n = 18). The animals were anesthetized by subcutaneous injection of xylazine (10 mg/kg) and ketamine (40 mg/kg). Once the ovaries were exposed, they were removed laparoscopically and the intact ovarian tissue was transplanted in a peritoneal pocket created in the lateral abdominal wall, or in the same location without anastomosis. In groups D and C, cyclosporine 5 mg/kg/day was administered postoperatively. Biopsies from the ovaries were taken 360 days postoperatively for histological evaluation. Blood samples for serum concentrations of FSH, LH E2, and progesterone (PROG) were measured, on day 0 (day of surgery), day 7, day 30, day 180, and day 360 postoperatively. The histological examination showed that the percentage of functional ovaries in group B was 68.18 %, in group C was 45 %, in group D was 63.16 %, and in group E was 38.89 %. No significant difference was found between groups B and C (p = 0.129), as well as between D and E (p = 0.14). The concentration of FSH and LH in group A was significantly higher than in the other groups (p 0.05). In group A, the concentrations of E2 and PROG were significantly lower than the other groups (p < 0.05), while statistically significant difference was observed between B, C, D, and E groups (p < 0.05). In conclusion, autologous and heterologous ovarian transplantation of intact ovaries without vascular anastomosis was performed successfully and the animal tolerance was excellent. Orthotopic and heterotopic grafts proved to be functional by means of ovarian function and histological examination.

Histomorphometric Evaluation of Superovulation Effect on Follicular Development after Autologous Ovarian Transplantation in Mice.

The effect of superovulation by pregnant mare serum gonadotropin (PMSG) on autologous transplanted ovaries in the lumbar muscles of mice was histomorphometrically evaluated using the indices of number and volume of different kind of follicles and volume of corpora lutea, ovary, and stroma. Angiogenesis was observed after mouse ovarian transplantation on days 14 and 21 after ovarian grafting. After transplantation, the total number and volume of primary and secondary follicles reduced, while PMSG superovulation increased the total number and total volume of tertiary follicles and also the ovarian volume after transplantation. Transplantation increased the average size of primary, secondary, and tertiary follicles. Therefore, primary and secondary follicles can survive after autologous transplantation but their reservations diminished by increasing the time of transplantation. However, number of tertiary follicles and their response to superovulation increased over time after transplantation.

N-acetylcysteine improves morphologic and functional aspects of ovarian grafts in rats.

To evaluate morphological and functional aspects of the ovarian graft in transplanted rats treated with NAC. Female Wistar rats, virgin, 3 to 4 months old, weighing 200-250 grams were used in experiments. The rats have been kept in proper sanitary conditions, receiving food and water ad libitum. Five groups (n=10, each) were constituted: 4 groups treated subcutaneously with NAC, at doses of 150, 300, 600 and 1200 mg/kg (NAC150, NAC300, NAC600 and NAC1200, respectively), one hour of before the ovarian transplantation and control group (GTx) - treated with physiological solution and submitted to ovarian transplantation. The rats were anesthetized and submitted to autologous left ovarian transplantation, without anastomosis in retroperitoneum, and contralateral oophorectomy. During follow-up of 4 or 15 days, the estrous cycle was evaluated by vaginal smears to determine cycle regularity. At the end of 4th or 15th days, rats were re-anesthetized and blood and graft were obtained to estradiol analysis and morphological assessment. Data were analysed by One Way Analysis of Variance (ANOVA) or ANOVA on ranks complemented by Student-Newman-Keuls test. At 4th day, viable follicles in the graft did not altered by NAC treatments. The NAC300 and NAC600 groups showed increasing in follicle atresia (p=0.012) compared to GTx and NAC1200 group. At 15th day, 50% of GTx, NAC150, and NAC300 rats showed regular oestrous cycle; 83% of NAC600 and 100% of NAC1200 rats returned to regular cycle. NAC1200 group showed increasing in primordial follicle compared to GTx, NAC150 or NAC300 (p=0.011). NAC did not interfere in estradiol levels after 4 or 15 days of transplantation. In autologous ovarian transplantation, high dose of NAC promotes graft viability with recovery of estrous cycle.

Autologous and allogeneic ovarian orthotopic transplantation: morphologic, endocrinologic and natural pregnancy assessment

To assess pregnancy of rabbits submitted to bilateral ovariectomy and orthotopic allogeneic or autologous intact and sliced ovarian transplantation without a vascular pedicle and to determine the morphofunctional aspects of the transplanted ovaries. Fifty-six female rabbits were studied. The ovaries were removed and orthotopically transplanted or replaced without vascular anastomoses: Group 1 (n=8), only laparotomy and laparorrhaphy, Group 2A (n=8) intact ovaries were transplanted on both sides, Group 2B (n=8) both ovaries were sliced and orthotopically transplanted, Group 2C (n=8), an intact ovary was transplanted on one side and a sliced ovary on the other side. In Group 3 the ovaries were reimplanted according to the procedure and subgroups described for Group 2. Three months later, the animals were paired with males for copulation. Estradiol, progesterone, FSH and LH hormone levels were assessed. A histologic study was carried out, and the number of pregnancies and litters were also determined. Chi-square test compared the number of pregnancies and litters. One-way ANOVA and the Tukey-Kramer tests compared the hormonal dosages. Pregnancies occurred in seven (87.5%) rabbits of Group 1, in three rabbits (37.5%) of Groups 2A and 3A, in four rabbits (50%) of groups 2B, 3B and 3C, and in five (62.5%) of group 2C. Normal hormone serum levels and histology confirmed the vitality of all ovaries. Orthotopic allogeneic and autologous ovarian transplantation without a vascular pedicle is viable in rabbits, and preserves their hormonal levels and fertile functions.

Improving autologous ovarian transplantation with frozen-banked tissue: robotic assistance and the utilization of a human regenerative matrix

OBJECTIVE: Ovarian transplantation (OTx) with frozen-banked ovarian tissue is evolving since the first report in 1999. One of the limitations of OTx is the large follicle loss due to initial ischemia. Our aim was to develop a new surgical technique to improve tissue vascularization and graft survival with the utilization of a human regenerative matrix (alloderm, AD) and robotically assisted surgery (DaVinci). DESIGN: Experimental. MATERIALS AND METHODS: Ovarian tissue (OT)/AD integration and neo-vascularization were studied in SCID mice using AD thicknesses 0.38-2.08 mm. GV oocytes were co-cultured with AD for 24h to test for ovo-toxicity. OTx was then simulated in porcine and inanimate models. The patient was a 36-year-old breast cancer survivor. Ovarian cortical biopsies were slow frozen before chemotherapy. Because the patient experienced ovarian failure, as confirmed by multiple FSH and LH levels > 40 mIU/mL, and she desired pregnancy, we performed OTx 2 years later. Larger pieces (2x2-2x4-mm) were sutured on AD microscopically, and then grafted onto L ovary using DaVinci. Small OT pieces (1x1-1x2-mm) were injected subcortically into the R ovary and the ovarian fossa, using a 14-gauge needle. Ovarian blood flow was measured by Doppler-ultrasound pre and post OTx. RESULTS: Optimal neo-vascularization, integration, and surgical handling were achieved with medium thickness (0.79-1.27 mm) AD (vascular density 27.0 ± 5.5 vs 15.5 ± 3.7 in controls, p<.001)., which was used for OTx. No ovo-toxicity was observed with AD (n=28, 89% survival) vs. none (n=26, 96% survival) (p=0.6). Compared to preop, ovarian blood flow significantly improved in the L (RI: from 0.8 to 0.5 and PSV 16.5 to 32.6 cm/s) but not the R ovary (RI: from 0.9 to 0.8, PSV 21 to 22 cm/s), 2 wk post op. Follow up by 4 weeks showed antral follicle development in both ovaries. CONCLUSION: We described for the first time a robotically assisted OTx involving the novel use of a regenerative matrix (ROBOTx), which may improve tissue revascularization and OTx outcomes.

Vessels arise in primate ovarian xenografts primarily from grafted ovarian tissue rather than from the mouse host, & neo-vascularization may increase with vascular endothelial growth factor (VEGF)

OBJECTIVE: Autologous ovarian tissue banking & transplantation may preserve fertility for women undergoing gonadotoxic chemotherapy. Because of the risk of cancer transmission within the autologously transplanted ovarian tissue, xenografting may be an alternative. The objectives of this study are to define whether vessels in xenografted ovarian tissue arise from the mouse host or the transplanted ovary, and whether VEGF increases neo-vascularization.

Allogeneic and autologous ovarian orthotopic transplantation without a vascular pedicle: Morphological, endocrinologic and natural pregnancy assessment

To assess the natural pregnancy of rabbits submitted to bilateral oophorectomy and orthotopic allogeneic or autologous intact and sliced ovarian transplantation without a vascular pedicle, and to determine the morpho-functional aspects of the transplanted ovaries. Fifty-six female rabbits were studied. The ovaries were removed and orthotopically transplanted or replaced without vascular anastomoses: Group 1 (n = 8), only laparotomy and laparorrhaphy; Group 2A (n = 8), intact ovaries were transplanted on both sides; Group 2B (n = 8), both ovaries were sliced and orthotopically transplanted; Group 2C (n = 8), an intact ovary was transplanted on one side and a sliced ovary on the other side; Groups 3A-C, the ovaries were reimplanted according to the procedure and subgroups described for Group 2. Three months later, the animals were paired with males for copulation. Estradiol, progesterone, FSH and LH hormone levels were assessed. A histological study was carried out, and the number of pregnancies and litters were also determined. Chi-squared test compared the number of pregnancies and litters. One-way anova and the Tukey-Kramer tests compared the hormonal dosages. Pregnancies occurred in seven rabbits of Group 1, in 37.5% of rabbits in Groups 2A and 3A, in 50% of groups 2B, 3B and 3C, and in 62.5% of group 2C. Hormone levels and histology confirmed the vitality of all ovaries. Intact or sliced orthotopic allogeneic and autologous ovarian transplantation without a vascular pedicle is viable in rabbits, and preserves their hormonal and fertile functions.

Avaliação pós-transplante de ovários íntegros e fatiados sem anastomose vascular

To assess the natural pregnancy and to determine the morphofunctional aspects of ovaries of rabbits submitted to bilateral oophorectomy and orthotopic allogeneic or autologous intact and sliced ovarian transplantation without a vascular pedicle. Fifty-six female New Zealand White and California rabbits were studied. The ovaries were removed and orthotopically transplanted or replaced without vascular anastomoses: Group 1 (n = 8), only laparotomy and laparorrhaphy were performed; Group 2A (n = 8) intact ovaries were reimplanted on both sides; Group 2B (n = 8) both ovaries were sliced and orthotopically reimplanted; Group 2C (n = 8), an intact ovary was reimplanted on one side and a sliced ovary on the other side; Group 3A (n = 8) intact ovaries were transplanted on both sides, Group 3B (n = 8) both ovaries were sliced and orthotopically transplanted, Group 3C (n = 8), an intact ovary was transplanted on one side and a sliced ovary on the other side. Three months later, the females were paired with males for copulation. Estradiol, progesterone, follicle stimulating hormone and luteinizing hormone levels were assessed. The morphological aspect of the ovaries was studied and the number of pregnancies and litters were also determined. The number of successful pregnancies and the number of litters was compared between the groups by the chi-square test. One-way ANOVA and the Tukey-Kramer tests compared the hormonal dosages. The significance was of p < 0.05. Pregnancies occurred in seven (87.5%) rabbits of Group 1, in 37.5% in Groups 2A and 3A, in 50% of groups 2B, 2C and 3B, and in 62.5% of group 3C. Hormone levels and histology confirmed the vitality of all ovaries. Intact or sliced orthotopic allogeneic and autologous ovarian transplantation without a vascular pedicle is viable in rabbits, and preserves their fertility and hormonal functions.