scholarly journals Establishment of the ovarian tissue cryopreservation protocol on bovine model

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 ◽  
...  
Keyword(s):  
Ovarian Function ◽  
Ovarian Tissue ◽  
Neutral Red ◽  
Slow Freezing ◽  
Ovarian Tissue Cryopreservation ◽  
Control Group ◽  
Ovarian Cortex ◽  
Group 2 ◽  
Tissue Cryopreservation ◽  
Cryopreservation Protocol

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.

scholarly journals The Effectiveness of Anti-Apoptotic Agents to Preserve Primordial Follicles and Prevent Tissue Damage during Ovarian Tissue Cryopreservation and Xenotransplantation

2021 ◽  
Vol 22 (5) ◽  
pp. 2534
Author(s):  
Sanghoon Lee ◽  
Hyun-Woong Cho ◽  
Boram Kim ◽  
Jae Kwan Lee ◽  
Tak Kim
Keyword(s):  
Tissue Damage ◽  
Ovarian Function ◽  
Ovarian Tissue ◽  
Slow Freezing ◽  
Ovarian Tissue Cryopreservation ◽  
Second Step ◽  
Control Group ◽  
Dna Breaks ◽  
Primordial Follicles ◽  
Tissue Cryopreservation

The purpose of this study is to investigate the effectiveness of sphingosine-1-phosphate (S1P) and Z-VAD-FMK (Z-VAD) as anti-apoptotic agents to preserve ovarian function and prevent tissue damage during ovarian tissue cryopreservation and transplantation. This study consisted of two steps, in vitro and in vivo. In the first step, human ovarian tissues were cryopreserved using slow-freezing media alone, S1P, or Z-VAD (control, S1P, Z-VAD group); based on the outcomes in these groups, Z-VAD was selected for subsequent xenotransplantation. In the second step, human frozen/thawed ovarian tissues were grafted into fifty mice divided into three groups: slow-freezing/thawing and transplantation without an anti-apoptotic agent (Trans-control) and xenotransplantation with or without Z-VAD injection (Trans-Z-VAD-positive and Trams-Z-VAD-negative groups, respectively). In the first step, the Z-VAD group had a significantly higher primordial follicular count than the S1P (p = 0.005) and control groups (p = 0.04). Transplanted ovarian tissues were obtained 4 weeks after transplantation (second step). Angiogenesis was significantly increased in the Z-VAD-negative (p = 0.03) and -positive (p = 0.04) groups compared to the control group. This study demonstrated that slow-freezing and transplantation with Z-VAD is an effective method for preserving primordial follicle counts, decreasing double-strand DNA breaks, and increasing angiogenesis in a mouse model. Further molecular and clinical studies are needed to confirm these results.

scholarly journals Ovarian tissue freezing and activation after thawing: an update

2021 ◽  
Vol 26 (1) ◽  
Author(s):  
Li-fan Peng
Keyword(s):  
Granulosa Cells ◽  
Clinical Decision Making ◽  
Ovarian Function ◽  
Ovarian Tissue ◽  
Clinical Decision ◽  
The Body ◽  
Endocrine Function ◽  
Ovarian Tissue Cryopreservation ◽  
Main Body ◽  
Tissue Cryopreservation

Abstract Background With the growth of women’s age, ovarian failure can be caused by various factors. For the women who need chemotherapy because of cancer factors, the preservation of fertility is more urgent. The treatment of cancer is also a process in which all tissues and organs of the body are severely damaged, especially in the reproductive system. Main body 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. Vitrification and slow freezing are two common techniques applied for ovarian tissue cryopreservation. Thus, cryopreserved/thawed ovarian tissue and transplantation act as an important method to preserve ovarian function during radiotherapy and chemotherapy, and ovarian cryopreservation by vitrification is a very effective and extensively used method to cryopreserve ovaries. The morphology of oocytes and granulosa cells and the structure of organelles were observed under the microscope of histology; the hormone content in the stratified culture medium of granulosa cells with the diameter of follicle was used to evaluate the development potential of ovarian tissue, and finally the ovarian tissue stimulation was determined by the technique of ovarian tissue transplantation. Conclusions Although there are some limitations, the team members still carry out this review to provide some references and suggestions for clinical decision-making and further clinical research.

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.

Mouse ovarian tissue vitrification on copper electron microscope grids versus slow freezing: a comparative ultrastructural study

2015 ◽  
Vol 27 (7) ◽  
pp. 1020 ◽  
Author(s):  
Ferda Topal-Celikkan ◽  
Sinan Ozkavukcu ◽  
Deniz Balci ◽  
Sibel Serin-Kilicoglu ◽  
Esra Atabenli-Erdemli
Keyword(s):  
Electron Microscopy ◽  
Electron Microscope ◽  
Cancer Therapy ◽  
Ovarian Tissue ◽  
Light And Electron Microscopy ◽  
Slow Freezing ◽  
Ovarian Tissue Cryopreservation ◽  
Control Group ◽  
Primordial Follicles ◽  
User Friendly

There are many reasons, including cancer therapy, for premature ovarian failure and infertility. Oocyte, embryo and ovarian cryopreservation are current options for fertility preservation. Ovarian tissue cryopreservation is essential in patients whose cancer therapy cannot be delayed, including prepubertal girls, and is mostly performed using slow freezing. In the present study, mouse ovarian tissues were vitrified on copper electron microscope grids (n = 18) or conventionally slow frozen (n = 18). Post-thaw tissues were examined histologically using light and electron microscopy and compared with the control group. According to light microscopy observations, antral follicles were found to be better preserved with the slow freezing technique rather than vitrification. Electron microscopy revealed swollen mitochondria in the oocyte cytoplasm, condensations in the zona pellucida, breakages in the junctions of granulosa cells and vacuolisation in the extracellular space in pathologic follicles, which were relatively more frequent, in the vitrification group after thawing. These results indicate that ovarian slow freezing is preferable than vitrification on copper electron microscope grids, especially for larger follicles. Conversely, vitrification of ovarian pieces using cooper grids is user-friendly and provided good protection for primordial follicles and stromal cells. There is a need for further studies into advanced tissue vitrification techniques and carriers.

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

scholarly journals FERTILITY PRESERVATION: Follicle reserve loss in ovarian tissue transplantation

Reproduction ◽  
2019 ◽  
Vol 158 (5) ◽  
pp. F35-F44 ◽  
Author(s):  
Hadassa Roness ◽  
Dror Meirow
Keyword(s):  
Ovarian Function ◽  
Ovarian Tissue ◽  
Young Patients ◽  
Ovarian Tissue Cryopreservation ◽  
Ischemic Damage ◽  
Pharmacological Agents ◽  
Ovarian Tissue Transplantation ◽  
After Cancer ◽  
Tissue Cryopreservation ◽  
Follicle Activation

Ovarian tissue cryopreservation and transplantation (OTCP-TP) has progressed over the past decade from a revolutionary experimental procedure to a well-accepted treatment in many centers for young patients with a high risk of ovarian failure after cancer treatment. The procedure is remarkably successful, with studies reporting return of ovarian function in up to 95% of graft recipients and pregnancy rates of between 30 and 50%. The most significant limitation of OTCP-TP is the massive loss of follicles that occurs following transplantation, which is primarily attributed to ischemic damage and follicle activation. We review the current approaches to reducing follicle loss and maximizing graft lifespan via pharmacological agents which reduce ischemic damage and follicle activation. We further discuss the value and disadvantage of inducing follicle activation in the graft as a means of generating mature follicles in the immediate short term.

scholarly journals FERTILITY PRESERVATION: Freezing of ovarian tissue and clinical opportunities

Reproduction ◽  
2019 ◽  
Vol 158 (5) ◽  
pp. F27-F34 ◽  
Author(s):  
C Yding Andersen ◽  
L S Mamsen ◽  
S G Kristensen
Keyword(s):  
Fertility Preservation ◽  
Ovarian Function ◽  
Ovarian Tissue ◽  
Genetic Diseases ◽  
Endocrine Function ◽  
Ovarian Tissue Cryopreservation ◽  
Ovarian Insufficiency ◽  
Technical Aspects ◽  
Increased Risk ◽  
Tissue Cryopreservation

Ovarian tissue cryopreservation (OTC) is mainly used for fertility preservation in girls and women facing a gonadotoxic treatment. If the woman subsequently becomes menopausal, the ovarian tissue may be transplanted to regain ovarian function, including fertility. The method was developed more than two decades ago and today thousands of women worldwide have undergone OTC. Fewer than 500 patients have had tissue transplanted and close to 100% of those regain ovarian function. Several technical aspects of OTC are now becoming more established, including high quantitative follicle survival, defining the size of the tissue resulting in optimal tissue revascularisation and follicle loss resulting from transport of ovarian tissue prior to freezing. We have used OTC to safeguard fertility in patients with genetic diseases, which for some diagnoses is purely experimental, as no transplantations is yet been performed. Usage of OTC beyond fertility is now also being considered; here, the endocrine function of follicles is the focus. It has been suggested that ovarian tissue stored in the reproductive years may be used to avoid premature ovarian insufficiency (POI) when there is a familial disposition or to postpone menopause in patients with an increased risk of osteoporosis or cardiovascular diseases. The benefit of OTC beyond fertility requires, however, actual clinical studies. The current review includes several recent technical aspects with contributions from Denmark building on some of the early work by Roger Gosden.

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