Daily blood samples were collected during the study period. Nacetylcysteine improves function and follicular survival in mice. Ovaries in vitrified groups were processed into thin slices then vitrified in cryotubes using vitrification medium and kept in liquid nitrogen for 21 days, rapidly thawed and transplanted into back muscles for 2 and 4 weeks. Pdf the effect of vitrification on follicular morphology of. Inthe present study, weconsideredthe hypothesis to. Vitrification of ovarian tissue is a useful method for preservation of female mouse germ cells and live pups following transplantation of vitrified warmed ovarian tissue 46. Oct 01, 2008 in this study, the efficiency of ovarian vitrification using ethylene glycol as a cryoprotectant was investigated by comparison of vitrified and nonvitrified samples by follicular viability assessment with trypan blue staining, transmission electron microscopy observation, the in situ tunel technique, and interanucleosomal dna fragmentation gel. Dec 26, 2014 the aim of this study was to evaluate the influence of two vitrification techniques on the extra cellular matrix ecm and ovarian follicular development. This study was designed to examine the effects of hp on invitro maturation of oocytes and cell death in cumulus cells derived from vitrified warmed mouse ovaries.
To date, there have been no studies which show the efficacy of embryo development until blastocyst stage from vitrified warmed oocytes compared to fresh oocytes from the same patients. Rescue of caprine fetal ovaries, vitrification and follicular. Human ovarian tissue biopsies from 23 women after caesarean section were transported to the laboratory within 2 hours, and then they were cut into small pieces. Oocyte vitrification versus ovarian cortex transplantation in fertility. Mem supplemented with 5% fetal bovine serum fbs for 7 days. In the laboratory, the ovarian medullary part was removed by a surgical blade, and the cortical tissue was cut into small pieces. Jan 01, 2020 the purpose of this study was to assess the viability and growth of follicles in ovarian tissues of cattle vitrified using two nonpermeating cryoprotectants sucrose and trehalose and two cryodevices cryovial and cryotop. Article successful vitrification of bovine and human. Objective this study was designed to evaluate the effects of vitrification and in vitro culture of human ovarian tissue on the expression of oocytic and follicular cellrelated genes. Gross morphology of bovine ovarian tissue vitrified using the cryotissue method. This prospective study aims to evaluate the feasibility of vitrifying in vitro matured oocytes ivm isolated at the time of ovarian tissue. Evaluation of the effects of pentoxifylline on stereological changes.
Our observation showed no significant difference between the mrna levels of some proapoptotic genes, including fasl, bax, p53, and caspase3 and antiapoptotic gene bcl2 in non vitrified and. Vitrification of in vitro matured oocytes collected from. Two methods of vitrification followed by in vitro culture. For each patient, the preservation status of the ovarian tissues cryopreserved using the two methods was compared with a fresh specimen, as well as between them using lm, tem and lscm. Ovarian tissue obtained from three of these women was only cultured for 24 h as non vitrified cultured controls. Pdf vitrification of zebrafish danio rerio ovarian follicles. Pdf effect of antifreeze protein on mouse ovarian tissue. Ovarian cryopreservation is a technique in artificial. Ultrastructural and morphalogical changes of mouse ovarian. Pdf the effect of vitrification on follicular morphology.
Vitrification and slow freezing are two common techniques applied for. Preservation of mouse ovarian tissue follicle morphology. In this study, we evaluated the incidence of apoptosis at the ultrastructural levels and expression of some apoptosisrelated genes in vitrified human ovarian tissue just after warming. Materials and methods in this experimental study, ovarian tissue samples were obtained from eight transsexual women. However, it has been shown that vitrification is often associated with ultrastructural damage of follicular cells and oocytes 711. Feb, 2020 considering that the overall survival rate of vitrified oocytes has previously been evaluated at 85. Effect of human ovarian tissue vitrificationwarming on. The aim of the present study was to assess the efficiency of nonequilibrium vitrification compared to conventional slow freezing for ovarian cortex cryopreservation. Effects of vitrification solutions and equilibration times on. Article successful vitrification of bovine and human ovarian. Morphological and molecular aspects of in vitro culture of.
In vitro survival of follicles in prepubertal ewe ovarian cortex. Short term culture of vitrified human ovarian cortical tissue to. Aim of this study, is to demonstrae an improved convetional vitrification method on mouse ovarian tissue using different concentrations of ethylene glycol eg andor dimetyl sulfoxide dmso and eg. In this experimental study, vitrified and non vitrified ovaries from 7 day old neonate. Effects of vitrification solutions and equilibration times. The accumulation of vitrified oocytes is a strategy to. Supplementation of culture media with vitamin e improves mouse. Pdf morphologic changes in fresh and vitrified mouse. In vitro growth and maturation as well as fertilization of.
Pieces of ovarian tissue of weekold japanese quail coturnix japonica were frozen at 0. This report describes a live birth using vitrified warmed oocytes and fertilitysparing surgery in a woman with invasive ovarian cancer stage ic. Effects of pentoxifylline on the histological and ultra. Autotransplantation of vitrified warmed ots after ivc 0 to 4 h using the eds or es protocol was performed, and the grafts were recovered after 3 weeks. Pdf vitrification of zebrafish danio rerio ovarian. Pdf live birth using vitrifiedwarmed oocytes in invasive. After various exposure times of cryoprotectant solution 5 min, 10 min, and 20 min, respectively, cryoprotectant. The purpose of this study was to assess the viability and growth of follicles in ovarian tissues of cattle vitrified using two nonpermeating cryoprotectants sucrose and trehalose and two cryodevices cryovial and cryotop. Stereological estimation of ovarian oocyte volume, surface. Vitrification of ovarian tissue the vitrification solution used in this study contained 40% ethylene glycol sigmaaldrich. Samples were cut into small fragments and were then assigned to vitrified and non vitrified. Assessment of longterm function of heterotopic transplants. Six hemiovaries from six ewes aged 6 to 12 months were vitrified. Samples were cut into small fragments and were then assigned to vitrified and non vitrified groups.
A second large group of patients were young women average age, 29 years with surplus oocytes after undergoing ivf, icsi, or ttomi tubal transfer of oocytes microinjected treatments. Vitrified bovine ovarian tissue was translucent in liquid nitrogen 196c. A sensitivity analysis revealed that no pregnancy occurred in the oct group when the ovarian tissue was harvested beyond 36 years old, while. Normal gestations and live births after orthotopic. Vitrified ovaries from neonate f1 hybrid mice, candidates for transplantation to treated or. The measured outcomes included the numbers of retrieved and vitrified oocytes, and direct medical costs. Production of donorderived offspring from cryopreserved. Based on results obtained from this stage, the vitrification ability of 24 combinations using. Angiogenesis and follicular development in ovarian tissue of. Pdf expression of folliculogenesisrelated genes in. Abstract the effect of nacetylcysteine nac on mouse ovary heterotopic. Nov 23, 2011 background in the past few years, cryopreservation of ovarian tissue has become an established procedure proposed in many centers around the world and transplantation has successfully resulted in fullterm pregnancies and deliveries in human. In order to find the optimal exposure time of cryoprotectant, we performed a comparison of vitrification versus slow freezing according to the degree of normal morphology and apoptosis of human ovarian follicles. Pdf revascularisation in human ovarian tissue after.
Pdf first successful vitrification of salmonid ovarian tissue. In this study, we demonstrated that human early preantral follicles consume oxygen. Vitrification and xenografting of human ovarian tissue. In the autografted salinetreated group, mice were injected with normal saline for 7 days. Effects of erythropoietin on ischemia, follicular survival, and ovarian. Angiogenesis and follicular development in ovarian tissue. The transparent glassy appearance during cooling and warming was used to identify vitrified solution. Ovarian follicles were assessed for morphology, apoptosis, proliferation and fsh level. Estimating human ovarian nongrowing follicle number. Stereology data were analyzed using the mannwhitney test using graph. Modulatory effect of gonadotropins on rats ovaries after nandrolone decanoate administration. Pdf successful ongoing pregnancies after vitrification.
The ovarian tissue was vitrified in accordance with previously described methods with some modifications. The aim of the present study was to characterize the morphological and ultrastractural of mouse ovarian tissue with different cryoprotectant solution. Clinical grade vitrification of human ovarian tissue. Hydrostatic pressure improves invitro maturation of oocytes. Thus, the objective of this study was to investigate firstly the effect of vitrification, and secondly lif supplementation as an antiapoptotic and survival factor on the follicular development and incidence of apoptosis after longterm culture of human ovarian tissue. Purpose to investigate the effect of antifreeze protein afp supplementation on ovarian vitrification and transplantation. In this experimental study, vitrified and non vitrified ovaries from 7 day old neonate female mice were cultured using alphaminimum essential medium. Optimal vitrification protocol for mouse ovarian tissue.
We isolated such follicles from nonvitrified and vitrified ovarian cortical tissue after longterm transportation. The vitrified ovarian sections were stored in liquid nitrogen for at least 2 months 172 42 mean sem days. Preservation of mouse ovarian tissue follicle morphology and. Morphology, surface area of ovaries and percentage of normal follicles were evaluated and compared in both groups.
Oxygen consumption rate of early preantral follicles from. Pdf follicular dynamics in neonate vitrified ovarian. In this study, to identify the best method of cryopreservation, the ovarian cortex retrieved from oncological patients was cryopreserved by sfrt and vw protocols. This study evaluates the effect of melatonin on follicular dynamics in neonate vitrified ovarian grafts. Oct 01, 2010 this study assessed the effects of vitrification solutions and equilibration times on morphology of cynomolgus ovarian tissues. The ovarian cortex was fragmented 9 mm 3 and divided into six groups, viz. Patients who experienced ovarian hyperstimulation syndrome ohss had the highest e 2 peak scores and the greatest number of vitrified oocytes per woman. Isolation and viability of vitrified brown trout salmo trutta ovarian germ cells and qualitative. However, additional study is needed to confirm this observation. Hydrostatic pressure improves invitro maturation of. Original article analysis of apoptosis in cultured human. For warming, straws were warmed in water at 35c for 10 s and the tissue cubes were expelled into 1 ml of h199 containing 20% sss and 0. Preovulatory follicles were harvested from non vitrified and vitrified warmed 68 weekold female nmri mouse ovaries and randomly.
The results showed that ovarian responses were adversely affected by the hot summer as indicated by a reduced number of cl, total ova. Doublein vitro maturation increases the number of vitrified. In brief, the ovaries were placed in an equilibrium solution containing 7. Normal gestations and live births after orthotopic autograft. From a practical standpoint, another important consideration in determining ovarian ngf count for studies investigating reproductive aging is whether. Pdf follicular dynamics in neonate vitrified ovarian grafts. The ovarian tissues were obtained from 5 or 6 weeks aged icr mouse. Merdassi et al21 in his study concluded that it is crucial to validate the freezing protocol in order to evaluate the success of xenoor autograft transplantation of frozenthawed ovarian fragments though, surgical technique. The study was approved by the ethics committee of the karolinska institutet, karolinska university hospital huddinge. In this study, the efficiency of ovarian vitrification using ethylene glycol as a cryoprotectant was investigated by comparison of vitrified and nonvitrified samples by follicular viability assessment with trypan blue staining, transmission electron microscopy observation, the in situ tunel technique, and interanucleosomal dna fragmentation gel. The main goal of this study was to compare developmental competence and oxidative status of vitrified. This study aimed to evaluate the expression of the genes related to folliculogenesis after vitrification of mouse ovarian tissues using a twostep in vitro culture.
Currently, the most common methods for evaluation of cryopreserved ovarian tissue are morph. Comparison of livebirth defects after lutealphase ovarian stimulation vs. Pdf successful ongoing pregnancies after vitrification of. Effect of plateletrich plasma prp on ovarian structures in. The usage of this kit was simpler and more convenient than the slowfreezing method, indicating that the method has potential benefits and advantages for clinical use. Some pieces were vitrified and warmed and the others were considered as non vitrified group control. The ocr increased with developmental stage but did not seem to be affected by patient age or transportation duration in this confined patient group.
Prediction of embryo survival and live birth rates after. There were 22 retrievals performed and 342 oocytes vitrified and warmed. This study aimed to evaluate the use of goat fetal ovarian tissue for. Find, read and cite all the research you need on researchgate. Therefore, the aims of the present study are to correlate the degree of expansion and embryo quality with survival and live birth in vitrified warmed blastocysts. Straws containing slowfrozen samples were thawed in ice water, and vitrified samples were removed from the vials and transferred. Human ovarian tissue vitrificationwarming has minor effect.
Nov 01, 2012 the aim of the present study was therefore to assess the morphology, survival, and development of preantral follicles from vitrified warmed human ovarian tissue using a vitrification protocol established by huang et al. Mar 01, 2004 in this study, mouse ovaries were vitrified using a new vitrification kit with a polyester sheet for storage in liquid nitrogen. Materials and methods in this experimental study, we researched a total of 182 ovaries from 4weekold icr mice. Two methods of vitrification followed by in vitro culture of. Surviving oocytes arrows of preantral follicles of vitrified thawed bovine ovarian. The equilibration solution included 20% ethylene glycol eg, and the vitrification solution included 40% eg, 18% ficoll, and 0. Pooled samples from different patients of various ages and reproductive backgrounds are often used in a single study, resulting in a lack of systematic comparison. The aim of this study was to evaluate the effects of preventive vitamin e.
Eleven patients aged 2041 years who underwent operative laparoscopy for benign ovarian cysts or cesarean section were enrolled in this study. In this experimental study, the gene expression of mmp2, mmp9, timp1, and timp2 in the isolated preantral follicles derived from fresh and vitrified ovaries of 1416 days old female mice through real time qrt pcr was evaluated. Human ovarian tissue cryopreservation has been successfully applied. This study evaluates the effect of ethylene glycole vitrification on follicular morphology of ovarian rat. Surviving oocytes arrows of preantral follicles of vitrified thawed bovine ovarian tissue hoechstpropidium iodide stain. Ovarian failure is diagnosed by ovarian destruction and reducing sex hormonal levels. Human ovarian tissue vitrificationwarming has minor. Morphological, ultrastructural and functional imaging of.
In the other part of this study, we evaluated, for the first time, the expression of some apoptosisrelated genes in vitrified human ovarian tissue. Comparison between slow freezing and vitrification for. Live offspring have been born from vitrified mouse ovarian preantral follicles matured in vitro, and orthotopic autografting of vitrified warmed sheep hemi. Pdf comparative study of results obtained through fresh.
Ovarian function and reproductive outcome after ovarian tissue. Research open access preservation of mouse ovarian. Human ovarian tissue samples were collected from five transsexual women. Morphological and functional preservation of preantral. Effect of human ovarian tissue vitrificationwarming on the. Inthe present study, weconsideredthe hypothesis to accu. The morphological study using hematoxylin and eosin and massons trichrome staining was done. Morphologic, ultrastructural, and biochemical identification. We conducted a prospective cohort study associated with a costeffectiveness analysis in a tertiarycare university hospital. The measured outcomes included the numbers of retrieved and vitrified. Ultrastructural analysis of vitrified rat ovarian tissue. Influence of the vitrification solution on the angiogenic.
Research open access preservation of mouse ovarian tissue. Case reporta 28yearold patient attended this study centre for a second opinion in january 2010. Eighty ovaries belonging to 40 rats are divided into 2 groups. The optimal protocol for ot vitrification should maintain follicular structure and provide a higher capacity to survive throughout the subsequent ivc.
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