Human Ovarian Cryopreservation: Vitrification Versus Slow Freezing From Histology To Gene Expression.

2021 ◽  
Author(s):  
Pauline Jaeger ◽  
Cyrielle Fournier ◽  
Claire Santamaria ◽  
Eloise Fraison ◽  
Nicolas Morel-Journel ◽  
...  
Keyword(s):  
Gene Expression ◽  
Ovarian Tissue ◽  
Slow Freezing ◽  
Ovarian Tissue Cryopreservation ◽  
Fresh Tissue ◽  
Stage Of Development ◽  
Expression Of Genes ◽  
Significant Difference ◽  
Tissue Cryopreservation ◽  
Follicular Density

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.

scholarly journals Comparison between Slow Freezing and Vitrification for Human Ovarian Tissue Cryopreservation and Xenotransplantation

2019 ◽  
Vol 20 (13) ◽  
pp. 3346 ◽  
Author(s):  
Sanghoon Lee ◽  
Ki-Jin Ryu ◽  
Boram Kim ◽  
Dahyeon Kang ◽  
Yoon Young Kim ◽  
...  
Keyword(s):  
Ovarian Tissue ◽  
Primordial Follicle ◽  
Research Unit ◽  
University Hospital ◽  
Slow Freezing ◽  
Ovarian Tissue Cryopreservation ◽  
Ki 67 ◽  
Immunodeficient Mice ◽  
Human Ovarian Tissue ◽  
Tissue Cryopreservation

Two methods for the cryopreservation of human ovarian tissue were compared using a xenotransplantation model to establish a safe and effective cryopreservation method. Ovarian tissues were obtained from women who underwent benign ovarian surgery in the gynecology research unit of a university hospital. The tissues were transplanted into 112 ovariectomized female severe combined immunodeficient mice 4 weeks after slow freezing or vitrification cryopreservation. Tissues were retrieved 4 weeks later. Primordial follicular counts decreased after cryopreservation and xenotransplantation, and were significantly higher in the slow freezing group than in the vitrification group (p < 0.001). Immunohistochemistry and TUNEL assay showed that the Ki-67 and CD31 markers of follicular proliferation and angiogenesis were higher in the slow freezing group (p < 0.001 and p = 0.006, respectively) and DNA damage was greater in the vitrification group (p < 0.001). Western blotting showed that vitrification increased cellular apoptosis. Anti-Müllerian hormone expression was low in transplanted samples subjected to both cryopreservation techniques. Electron microscopy revealed primordial follicle deformation in the vitrification group. Slow freezing for ovarian tissue cryopreservation is superior to vitrification in terms of follicle survival and growth after xenotransplantation. These results will be useful for fertility preservation in female cancer patients.

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 ◽  
...  
Keyword(s):  
Subcutaneous Tissue ◽  
Ovarian Tissue ◽  
Fibrous Tissue ◽  
Ovarian Tissue Cryopreservation ◽  
Post Transplantation ◽  
Fresh Tissue ◽  
Tissue Samples ◽  
Antral Follicles ◽  
Ovarian Cortex ◽  
Tissue Cryopreservation

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.

Application of computed tomography to ovarian tissue cryopreservation by slow freezing

Cryobiology ◽  
2015 ◽  
Vol 71 (3) ◽  
pp. 546
Author(s):  
Ariadna Corral ◽  
Marcin Balcerzyk ◽  
Ángel Parrado ◽  
Christiani Amorim ◽  
Marie-Madeleine Dolmans ◽  
...  
Keyword(s):  
Computed Tomography ◽  
Ovarian Tissue ◽  
Slow Freezing ◽  
Ovarian Tissue Cryopreservation ◽  
Tissue Cryopreservation

Slow-freezing versus vitrification for human ovarian tissue cryopreservation

2014 ◽  
Vol 291 (2) ◽  
pp. 419-426 ◽  
Author(s):  
Silke Klocke ◽  
Nana Bündgen ◽  
Frank Köster ◽  
Ursula Eichenlaub-Ritter ◽  
Georg Griesinger
Keyword(s):  
Ovarian Tissue ◽  
Slow Freezing ◽  
Ovarian Tissue Cryopreservation ◽  
Human Ovarian Tissue ◽  
Tissue Cryopreservation ◽  
Human Ovarian Tissue Cryopreservation

scholarly journals Ovarian Tissue Reservation and Risk of Reimplantation in a Young Girl with Acute Lymphocytic Leukemia after 6-Month Chemotherapy: A Case Report

2021 ◽  
Author(s):  
Mojgan Karimi-Zarchi ◽  
Mohammad Ali Khalili ◽  
Fariba Binesh ◽  
Vatanparast Mahboubeh
Keyword(s):  
Fertility Preservation ◽  
Acute Lymphocytic Leukemia ◽  
Ovarian Tissue ◽  
Treatment Protocol ◽  
Lymphocytic Leukemia ◽  
Ovarian Tissue Cryopreservation ◽  
Normal Range ◽  
History Of ◽  
Tissue Cryopreservation ◽  
Follicular Density

AbstractIn the fertility preservation programs, ovarian cryopreservation is usually offered when the risk of premature ovarian failure is high (>30–50%) while the risk of ovarian metastasis is low. According to the guidelines, it must be done before the patient receives chemotherapy. A 22-year-old girl with acute lymphocytic leukemia was a candidate for ovarian cryopreservation after 6 months of chemotherapy. Despite chemotherapy, the anti-Mullerian hormone survey was within normal range. Ovarian tissue cryopreservation was done. In the histology survey, follicular density was 7.48. This case shows that only having a history of chemotherapy does not exclude the patient from the fertility preservation program. Regarding referring the patients for fertility preservation, cumulative factors such as a history of gonadotoxic treatment, age, and treatment protocol should be considered. In addition, the case was negative for assessing of CD45 marker. New data may challenge previous strict criteria, and extend the indications of this effective method in preserving fertility among cancer patients.

Improving ovarian tissue cryopreservation for oncologic patients: slow freezing versus vitrification, effect of different procedures and devices

2014 ◽  
Vol 101 (3) ◽  
pp. 775-784.e1 ◽  
Author(s):  
Sonia Herraiz ◽  
Edurne Novella-Maestre ◽  
Beatriz Rodríguez ◽  
César Díaz ◽  
María Sánchez-Serrano ◽  
...  
Keyword(s):  
Ovarian Tissue ◽  
Slow Freezing ◽  
Ovarian Tissue Cryopreservation ◽  
Oncologic Patients ◽  
Tissue Cryopreservation
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