scholarly journals Function of Cryopreserved Cat Ovarian Tissue after Autotransplantation

Animals ◽  
2019 ◽  
Vol 9 (12) ◽  
pp. 1065 ◽  
Author(s):  
Janice M. V. Vilela ◽  
Ellen C. R. Leonel ◽  
Liudimila P. Gonçalves ◽  
Raísa E. G. Paiva ◽  
Rodrigo S. Amaral ◽  
...  

The aim of this study was to assess a slow-freezing protocol of cat ovarian tissue cryopreservation using autotransplantation. Four adult queens were ovariohysterectomized and the ovaries were fragmented and cryopreserved. After one week, the grafts were thawed and autografted to the subcutaneous tissue of the dorsal neck of each queen, then randomly removed after 7, 14, 28, 49, and 63 days after transplantation. Percentages of morphologically normal primordial and growing follicles (MNFs) were 88% and 97%, respectively, in fresh tissue samples (fresh controls), and 74% and 100%, respectively, immediately after thawing (cryo D0). No MNFs were found after 49 days of transplantation. In both fresh control and cryo D0 fragments, granulosa cells were frequently in proliferation. Two morphologically normal antral follicles were detected in one queen on Day 28 post-transplantation. Connective tissue fibers increased, suggesting replacement of active ovarian cortex by fibrous tissue. Tissue vascularization was observed at 7 days after grafting, and wide blood vessels were clearly visible on Days 49 and 63. In conclusion, although follicular survival was low after cryopreservation and grafting of cat ovarian tissue, follicles were able to develop up to the antral stage, which is an encouraging outcome.

Reproduction ◽  
2011 ◽  
Vol 141 (2) ◽  
pp. 183-191 ◽  
Author(s):  
Xiaoqian Wang ◽  
Sally Catt ◽  
Mulyoto Pangestu ◽  
Peter Temple-Smith

Cryopreservation of ovarian tissue is an important option for preserving the fertility of cancer patients undergoing chemotherapy and radiotherapy. In this study, we examined the viability and function of oocytes derivedin vitrofrom pre-antral follicles as an alternative method for restoring fertility. Pre-antral follicles (specified as secondary follicle with a diameter around 100–130 μm) were mechanically isolated from vitrified-warmed and fresh adult mouse ovarian tissues and cultured for 12 days followed by an ovulation induction protocol at the end of this period to initiate oocyte maturation. Oocytes were then released from these follicles, fertilizedin vitro, and cultured to the blastocyst stage and vitrified. After storage in liquid nitrogen for 2 weeks, groups of vitrified blastocysts were warmed and transferred into pseudo-pregnant recipient females. Although most of the isolated mouse pre-antral follicles from fresh (79.4%) and vitrified (75.0%) ovarian tissues survived the 12-dayin vitroculture period, significantly fewer mature oocytes developed from vitrified-warmed pre-antral follicles than from the fresh controls (62.2 vs 86.4%,P<0.05). No difference was observed in embryo cleavage rates between these two groups, but the proportion of embryos that developed into blastocysts in the vitrification group was only half that of the controls (24.2 vs 47.2%,P<0.05). Nevertheless, live births of healthy normal pups were achieved after transfer of vitrified blastocysts derived from both experimental groups. This study shows that successful production of healthy offspring using anin vitrofollicle culture system is feasible, and suggests that this procedure could be used in cancer patients who wish to preserve their fertility using ovarian tissue cryopreservation.


2020 ◽  
Author(s):  
M S Zemyarska ◽  
B D Bjarkadottir ◽  
X Wei ◽  
C A Walker ◽  
S M Lane ◽  
...  

AbstractBackgroundOvarian tissue cryopreservation (OTC) is important for fertility preservation and conservation. Delay in OTC may be required for transport or workflow management, however little is understood about the effect of processing delay on the tissue.ObjectiveTo determine whether a delay of 24-48 hours to OTC affects primordial follicle (PF) health.MethodsOvaries (n=6 sheep) were processed immediately or after storage at 4°C (24h, 48h). Tissue was fixed fresh, after cryopreservation or 10-day xenotransplantation. Morphological assessment of follicle health and development was performed.FindingsA total of 1541 follicles were analysed. A 24h processing delay did not impact PF health in fresh or cryopreserved tissue. In fresh tissue a 48h delay had an adverse effect on follicle health (OR=2.47, 95% CI 1.29-4.71). Interestingly, a 48h delay resulted in cryopreserved tissue being less likely to be graded as unhealthy compared to control (OR=0.56, 95% CI 0.36-0.87). There was no difference in PF health or development across groups following xenotransplantation.ConclusionOvarian tissue can be stored for up to 48 hours prior to cryopreservation with no net impact on PF health.


2021 ◽  
Author(s):  
Thu Yen Thi Nguyen ◽  
Luciana Cacciottola ◽  
Alessandra Camboni ◽  
Joachim Ravau ◽  
Michel De Vos ◽  
...  

Abstract STUDY QUESTION Is there a possibility of reseeding cancer cells potentially present in frozen ovarian tissue from patients with central nervous system (CNS) tumours? SUMMARY ANSWER Malignancy reseeding in cryopreserved ovarian tissue from 20 patients with CNS tumours was not detected by histology, immunohistochemistry (IHC), molecular biology or xenotransplantation. WHAT IS KNOWN ALREADY Ovarian metastasis potential has been documented in patients with leukaemia, borderline ovarian tumours, advanced breast cancer and Ewing sarcoma. However, data on the safety of transplanting frozen-thawed ovarian tissue from cancer patients with CNS tumours are still lacking. STUDY DESIGN, SIZE, DURATION This prospective experimental study was conducted in an academic gynaecology research laboratory using cryopreserved ovarian cortex from 20 patients suffering from CNS tumours. Long-term (5 months) xenografting was performed in immunodeficient mice. PARTICIPANTS/MATERIALS, SETTING, METHODS Subjects enrolled in the study were suffering from one of six types of CNS tumours including medulloblastoma, ependymoma, primitive neuroectodermal tumours, astrocytoma, glioblastoma and germinoma. The presence of malignant cells was investigated with disease-specific markers for each patient in cryopreserved and xenografted ovarian tissue by histology, IHC via expression of neuron-specific enolase (NSE) and glial fibrillary acidic protein (GFAP), and reverse transcription droplet digital polymerase chain reaction (RT-ddPCR) for quantification of GFAP and ENO2 gene amplification. MAIN RESULTS AND THE ROLE OF CHANCE Serial sections of cryopreserved and xenografted ovarian tissue from 20 patients showed no malignant cells by histology. All samples were negative for NSE and GFAP, although these neural markers were expressed extensively in the patients’ primary tumours. Analysis by RT-ddPCR revealed no cancer cells detected in cryopreserved and xenografted ovarian fragments from subjects with astrocytoma, ependymoma, glioblastoma or medulloblastoma. Taken together, the study found no evidence of malignancy seeding in frozen-thawed and xenotransplanted ovarian tissue from patients affected by CNS cancers. LIMITATIONS, REASONS FOR CAUTION This analysis cannot guarantee complete elimination of disseminated disease from all cryopreserved ovarian cortex, since we are unable to examine the fragments used for transplantation. WIDER IMPLICATIONS OF THE FINDINGS This is the first study to be conducted in patients with CNS cancers undergoing ovarian tissue cryopreservation and transplantation, and clearly demonstrates no tumour seeding in their frozen-thawed and xenografted tissue. This information is vital for doctors to provide patients with meaningful and accurate advice on the possibilities and risks of ovarian tissue reimplantation. STUDY FUNDING/COMPETING INTEREST(S) This study was supported by grants from the Fonds National de la Recherche Scientifique de Belgique–the Excellence of Science (FNRS–EOS), number 30443682 awarded to M.-M.D. and T.Y.T.N., FNRS grant number 5/4/150/5 and FNRS-PDR Convention grant number T.0077.14 awarded to M.-M.D., grant 2018-042 from the Foundation Against Cancer awarded to A.C., and private donations (Ferrero, de Spoelberch). The authors declare no competing financial interests. TRIAL REGISTRATION NUMBER N/A.


2011 ◽  
Vol 9 (1) ◽  
pp. 150 ◽  
Author(s):  
Giovanna Fasano ◽  
Federica Moffa ◽  
Julie Dechène ◽  
Yvon Englert ◽  
Isabelle Demeestere

2021 ◽  
Author(s):  
Jennifer A. Shannon ◽  
Aishwarya Sundaresan ◽  
Orhan Bukulmez ◽  
Zexu Jiao ◽  
Sarah Capelouto ◽  
...  

AbstractOvarian tissue cryopreservation and subsequent autologous transplantation has allowed resumption of endocrine function as well as fertility in certain populations. However, graft function is short-lived due to ischemia and aberrant follicular activation post-transplantation. While many studies have focused on gene expression, we wanted to determine whether cryopreservation itself had a deleterious effect on regulatory elements that might influence transcriptional integrity and graft performance. In this study, we used Omni-ATAC to assess genome-wide chromatin accessibility in primary human follicles before and after cryopreservation. Omni-ATAC from fresh ovarian follicles identified active regulatory elements expected to be functional in oocytes and granulosa cells, and gene ontology was consistent with RNA translation/processing and DNA repair. While promoter accessibility was largely maintained in cryopreserved ovarian follicles, we observed a widespread increase in the number of accessible enhancers. Transcription factor motif analysis and gene ontology suggested that this dysregulation was focused around the epithelial-mesenchymal transition. Indeed, transcription factor binding was noted in major pathways involved in this transition: TGF-β and Wnt signaling. Overall, our work provides the first genomic analysis of active regulatory elements in matched fresh and cryopreserved ovarian follicles as they undergo the process of ovarian tissue cryopreservation. Our data suggest that the process of cryopreservation activates an epithelial-mesenchymal transition state, which may lead to graft burn-out post-transplantation. Optimizing this technique in relation to this transition may therefore be an important step towards improving graft longevity and patient outcomes in fertility preservation.Summary sentenceCryopreservation of ovarian cortical tissue results in activation of differentiation and EMT pathways in follicles, which may explain graft burnout after autotransplantation.


2016 ◽  
Vol 19 (2) ◽  
pp. 24-32
Author(s):  
Dung Thi Phuong Nguyen ◽  
Lan Thi Thu Nguyen ◽  
Quang Nhat Nguyen ◽  
Tuong Manh Ho ◽  
Loc Minh Tai Nguyen ◽  
...  

Ovarian tissue cryopreservation is a suitable method for fertility preservation on women receiving treatment that may threaten the ovarian function and subsequent fertility. The whole ovarian or a part of ovarian can be cryopreserved for future use. This study was aimed to establish ovarian tissue cryopreservation protocols on bovine model for human application in Vietnam. In this method, bovine ovarians were collected from a slaughterhouse and kept at 4 oC up to a maximum of 12 hours before doing experiments. The ovarian cortex was cut into pieces of 10x10x1 mm. These pieces were randomly divided into 3 groups: (1) fresh species (control group), (2) species were freezed by slow-freezing method and (3) pieces were freezed by vitrification. After thawing, ovarian cortex pieces were treated with Collagenase Ia for the follicle isolation. The isolated follicles then were stained with Neutral Red. The rate of viable follicles was used as the outcome measure to assess the efficiency of the cryopreservation protocol. In results, the rates of viable follicles were 72.46 ± 6.11 % and 59.09 ± 7.08 % after slow-freezing and vitrification comparing to the control group, respectively. This was the first study which successfully established a protocol of ovarian tissue cryopreservation on bovine model in Vietnam. The protocol should be improved for further application to human treatment in the near future.


2021 ◽  
Author(s):  
Pauline Jaeger ◽  
Cyrielle Fournier ◽  
Claire Santamaria ◽  
Eloise Fraison ◽  
Nicolas Morel-Journel ◽  
...  

Abstract Background: Cryopreservation of ovarian tissue is one of the strategies offered to girls and women needing gonadotoxic treatment, as a means of preserving their fertility. There are two methods of ovarian tissue cryopreservation: slow freezing, the reference method, and vitrification, an alternative method. The aim of the present study was to evaluate which of the two is the best method for human ovarian tissue cryopreservation. Each ovary was divided into 3 groups: Fresh, Slow freezing and Vitrification. In each group a histological study to evaluate follicular density and quality; and an evaluation of 6 gene expression (CYP11A, STAR, GDF9, ZP3, CDK2, CDKN1A) were performed. Results: We observed no significant difference in follicular density within these 3 groups. Slow freezing altered the pool of primordial follicles compared to the Fresh tissue (31.8% vs 55.9%, p = 0.046, respectively). The expression of genes involved in steroidogenesis varied after cryopreservation compared to the fresh group; CYP11A was under-expressed in both freezing groups compared to the fresh group and significantly under-expressed in the slow freezing group (p = 0.01), STAR was over-expressed in the slow freezing group and significantly under-expressed in the vitrification group (p = 0.01). Regarding the expression of genes involved in cell cycle regulation, CDKN1A was significantly under-expressed in both freezing groups (slow freezing: p = 0.0008; vitrification: p = 0.03) compared to the fresh group. Conclusion: Vitrification had no effect on the histological quality of the follicles at any stage of development compared to Fresh tissue. There was no significant difference in gene expression between the two techniques.


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