Immature Oocytes Research Articles

ASSESSING THE EFFICACY OF LONG-PROTOCOL OVULATION STIMULATION USING GONADOTROPIN-RELEASING HORMONE AGONIST IN WOMEN EXPERIENCING INFERTILITY AND COMORBIDITY OF LEIOMYOMA AND CHRONIC ENDOMETRITIS

Introduction. Nowadays infertility is one of the global medical and social issues worldwide. Assisted reproductive technologies, including in vitro fertilization, is one of the main techniques for restoring fertility. Among the main stages of IVF are controlled ovarian stimulation (COS), the optimal method of which is chosen depending on a numerous factor, such as the patient's age, the ovarian reserve, and the presence of concomitant gynecological or extragenital pathology. The prevalence of various combinations of gynecological pathologies, for example, the comorbidity of uterine leiomyoma and chronic endometritis, can affect the results of IVF in different ways, so scientists around the world are increasingly studying the management of such patients. The aim of the study is to compare controlled ovarian stimulation long-protocol parameters and its effectiveness in the therapy of infertile patients with the comorbidity of uterine leiomyoma and chronic endometritis. Methods and participants. We performed retrospective comparative analysis of anamnestic data of 130 women with infertility who underwent infertility treatment with assisted reproductive technologies. The patients were divided into three groups. In all groups, stimulation of superovulation was carried out following a long protocol with a GnRH agonist; p<0.05 was considered statistically significant during the statistical data processing. Results. The study has revealed that higher starting dose of gonadotropins, as well as higher total gonadotropin consumptions are required for the patients of the second study group with uterine leiomyoma and chronic endometritis, compared to the first study group and the control group. The number of retrieved oocytes, including mature ones, were the lowest as well as the number of immature, deeply immature and degenerative oocytes were the highest in the second study group. The lowest rate of normal fertilization (2pn) and, as a result, a lower blastocyst yield is recorded in the same group. The highest frequency of aborted pregnancies among achieved clinical pregnancies is observed in the group of infertile women with uterine leiomyoma and chronic endometritis. Conclusions. The presence of the uterine cavity pathology combined with persistent chronic inflammation of the uterine mucosa, as well as the absence of proper preparation for IVF-ET cycles are the main reasons for the negative infertility treatment outcomes in such patients. This justifies the need for appropriate optimal preparation and elimination of the identified negative factors before or during the infertility treatment.

Contribution of Immature Oocytes to the Potential of a Stimulation Cycle

Contribution of Immature Oocytes to the Potential of a Stimulation Cycle

EFFECT OF CRYOPROTECTANTS AND CRYOPRESERVATION METHODS ON THE QUALITY OF BUFFALO OOCYTES

This study was conducted to evaluate the effect of two types of cryoprotectants and two cryopreservation methods on viability and normal morphology of the immature and in vitro mature of post-thawed buffalo oocytes. The in vitro mature and immature oocytes were kept in the vitrification solution (VS1) for 5 min and the vitrification solution (VS2) for 15 sec. The results revealed that the proportion of buffalo oocytes found to be viable and normal morphology were significantly (P< 0.01) higher in using glycerol which is 76.14% and 55.1%; respectively. We conclude from the present study that the viability and normal morphology of post-thawed buffalo vitrified in vitro matured oocytes were significantly (P < 0.01) higher in glycerol than those obtained in dimethyl sulfoxide (DMSO) and in vitrification technique than rapid technique.

EFFECT OF ADDING ALOE VERA EXTRACT ON THE QUALITY OF BUFFALO OOCYTES USING TWO CRYOPRESERVATION METHODS

This study was conducted to evaluate the impact of two types of cryopreservation and different concentrations of Aloe vera as an antioxidant (0,10% and 20%) on the viability and normal morphology of immature and in vitro mature buffalo cryopreserved oocytes. The in vitro mature and immature oocytes were kept in cryopreservation solutions for rapid and vitrification techniques. The results revealed that the proportion of buffalo oocytes found to be viable and normal morphology were significantly (P<0.01) higher produced when using a high concentration (20%) of aloe vera on in vitro mature buffalo oocyte which is79.11% and 59.02%; respectively. We conclude from the present study that the viability and normal morphology of post-thawed buffalo cryopreserved in vitro matured oocytes were significantly (P < 0.01) higher produced in using a high concentration of aloe vera than other concentrations of aloe vera, from the side while with other side the vitrification technique than a rapid technique.

Comparison of luteal phase and follicular phase in-vitro maturation in women with oocyte maturation abnormalities

Research Question: Are there differences in immature oocyte retrieval following luteal phase in vitro maturation (IVM) compared to follicular phase IVM in women with oocyte maturation abnormalities (OMAs)?Design: From January 2019 to May 2023, a retrospective cohort study at a private IVF center included 32 women with 53 IVM cycles in Group 1 (follicular phase) and 19 women with 32 IVM cycles in Group 2 (luteal phase). Additionally, nine women had both follicular and luteal phase IVM cycles for intracycle variability analysis.Result(s): The groups had no differences in oocyte maturation stages at collection (p>0.05). The Group 1 and Group 2 exhibited comparable median MII oocyte rates per patient after 48 hours after the collection (40.0%, IQR: 0.0-66.7% vs. 22.5%, IQR: 0.0-52.9%) (p>0.05). The median fertilization rate in Group 1 (66.7%, IQR: 50.0-66.7%) was found to be comparable to that in the Group 2 (66.7%, IQR: 50.0-66.7%) (p>0.05). There were no significant differences between the groups in the yielded embryo's grades and pregnancy rates (p>0.05). Comparing follicular and luteal phase IVM within the same menstrual cycle in nine patients, no differences were observed in oocyte maturation rates (p>0.05).Conclusion(s): This study indicates that there are no significant differences in oocyte maturation, fertilization rates, embryo quality, or pregnancy outcomes between luteal phase and follicular phase IVM in women with OMAs. These findings suggest that luteal phase IVM can be used similarly to follicular phase IVM, offering a potential avenue to enhance embryo yield for women with OMAs.

C-type natriuretic peptide improves maternally aged oocytes quality by inhibiting excessive PINK1/Parkin-mediated mitophagy.

The overall oocyte quality declines with aging, and this effect is strongly associated with a higher reactive oxygen species (ROS) level and the resultant oxidative damage. C-type natriuretic peptide (CNP) is a well-characterized physiological meiotic inhibitor that has been successfully used to improve immature oocyte quality during in vitro maturation. However, the underlying roles of CNP in maternally aged oocytes have not been reported. Here, we found that the age-related reduction in the serum CNP concentration was highly correlated with decreased oocyte quality. Treatment with exogenous CNP promoted follicle growth and ovulation in aged mice and enhanced meiotic competency and fertilization ability. Interestingly, the cytoplasmic maturation of aged oocytes was thoroughly improved by CNP treatment, as assessed by spindle/chromosome morphology and redistribution of organelles (mitochondria, the endoplasmic reticulum, cortical granules, and the Golgi apparatus). CNP treatment also ameliorated DNA damage and apoptosis caused by ROS accumulation in aged oocytes. Importantly, oocyte RNA-seq revealed that the beneficial effect of CNP on aged oocytes was mediated by restoration of mitochondrial oxidative phosphorylation, eliminating excessive mitophagy. CNP reversed the defective phenotypes in aged oocytes by alleviating oxidative damage and suppressing excessive PINK1/Parkin-mediated mitophagy. Mechanistically, CNP functioned as a cAMP/PKA pathway modulator to decrease PINK1 stability and inhibit Parkin recruitment. In summary, our results demonstrated that CNP supplementation constitutes an alternative therapeutic approach for advanced maternal age-related oocyte deterioration and may improve the overall success rates of clinically assisted reproduction in older women.

C-type natriuretic peptide improves maternally aged oocytes quality by inhibiting excessive PINK1/Parkin-mediated mitophagy

The overall oocyte quality declines with aging, and this effect is strongly associated with a higher reactive oxygen species (ROS) level and the resultant oxidative damage. C-type natriuretic peptide (CNP) is a well-characterized physiological meiotic inhibitor that has been successfully used to improve immature oocyte quality during in vitro maturation. However, the underlying roles of CNP in maternally aged oocytes have not been reported. Here, we found that the age-related reduction in the serum CNP concentration was highly correlated with decreased oocyte quality. Treatment with exogenous CNP promoted follicle growth and ovulation in aged mice and enhanced meiotic competency and fertilization ability. Interestingly, the cytoplasmic maturation of aged oocytes was thoroughly improved by CNP treatment, as assessed by spindle/chromosome morphology and redistribution of organelles (mitochondria, the endoplasmic reticulum, cortical granules, and the Golgi apparatus). CNP treatment also ameliorated DNA damage and apoptosis caused by ROS accumulation in aged oocytes. Importantly, oocyte RNA-seq revealed that the beneficial effect of CNP on aged oocytes was mediated by restoration of mitochondrial oxidative phosphorylation, eliminating excessive mitophagy. CNP reversed the defective phenotypes in aged oocytes by alleviating oxidative damage and suppressing excessive PINK1/Parkin-mediated mitophagy. Mechanistically, CNP functioned as a cAMP/PKA pathway modulator to decrease PINK1 stability and inhibit Parkin recruitment. In summary, our results demonstrated that CNP supplementation constitutes an alternative therapeutic approach for advanced maternal age-related oocyte deterioration and may improve the overall success rates of clinically assisted reproduction in older women.

In Vitro Fertilization of Immature Bovine Oocytes Matured in a Protein-Free Media and Their Subsequent Development to the Blastocysts

Henüz doğmamış buzağılardan elde edilen fötal buzağı serumu, buzağının acı çekmesine neden olmaktadır. Hayvanlara daha az acı çektirilmesi ve hayvan refahı gibi konularda süregelen tartışmalar neticesinde özellikle son yıllarda, in vitro embriyo ve hücre kültüründe fötal serum ikamelerinin kullanılma olanaklarına dair çok sayıda araştırma yapılmaktadır. Bununla birlikte serum içerisinde mevcut olan ve konsatrasyonu tam olarak belirlenemeyen moleküllerin hücresel işlevler üzerine etkilerinden dolayı özellikle tekli moleküler yolakların araştırıldığı çalışmalarda serum kullanımı bazı çakışmalara neden olabilmektedir. Bu amaçla, serum ikamelerinin kullanım olanaklarının araştırıldığı bu çalışmada, in vitro matürasyon için knockout serumTM kullanımının ilk bölünme ve blastosiste kadar gelişen embriyo sayısının önemli oranda artırdığı tespit edilmiştir. Bununla birlikte sentetik serum ikamesi ve serum ikamesi 2 kullanımının ise elde edilen embriyo sayılarını bir miktar düşürdüğü belirlenmiştir.

Oocyte in-vitro maturation primed with letrozole-HCG versus FSH-HCG in women with oocyte maturation abnormalities: a retrospective study

Research QuestionThe objective of this study was to compare letrozole-hCG primed IVM with FSH-hCG primed IVM in women with oocyte maturation abnormalities (OMAs) defined as at least two failed in vitro fertilization (IVF) cycles where immature oocytes were retrieved. DesignThis retrospective study was conducted at a private fertility clinic from January 2009 to April 2023. The final analysis included 75 women in Group 1 (FSH-hCG primed IVM) and 52 women in Group 2 (letrozole-hCG IVM). Result(s)Group 1 demonstrated a significantly higher median number of oocytes obtained [9, (IQR: 1-5) vs. 5 (IQR: 1-18)] (p<0.001). However, the groups had no differences in oocyte maturation stages at collection (p>0.05). At the end of IVM, group 1 had 73/666 mature oocytes, while group 2 had 106/322 mature oocytes, and the median MII oocyte rate per patient was higher in group 2 [33.3%, (IQR: 66.7-100.0%) vs. 0.0%, (IQR: 0.0-22.2%)] (p<0.001). Moreover, Group 2 demonstrated a higher median fertilization rate [66.7%, (IQR: 50.0-100.0%) vs. 50.0%, (IQR: 0.0-66.7%)] (p=0.027). The group 2 had a higher proportion of Grade 1 (17.1% vs. 0%) and Grade 2 (58.5% vs. 6.3%) embryos (p<0.001). Notably, all pregnancies obtained in the study were in Group 2 (5 vs. 0; p=0.042). Conclusion(s)IVM with Letrozole-hCG priming in women with prior failed IVF cycles due to OMAs may result in mature oocytes, clinical pregnancies, and live births. The effectiveness of letrozole priming IVM on the subtypes of OMAs need further investigation with studies including higher number of cases.

Fertility Preservation Options for Transgender Patients: An Overview

Fertility preservation technologies have existed for decades, and the field is rapidly advancing; limited data exist regarding the use of these technologies by transgender patients. Many options are available for transgender patients who wish to preserve fertility before transitioning. These options include the cryopreservation of gametes, embryos, or ovarian tissue. Currently, ejaculated, or testicular sperm, immature oocytes, and ovarian tissue can be preserved for later use, but no such use option exists for immature testicular tissue. Many financial, sociological, and legal barriers and a lack of awareness among physicians and patients also hinders the utilization of these fertility preservation services. While options are abundant, usage rates are relatively low. The initial data regarding the successful use of preserved tissues appears promising, with birth rates not dissimilar to non-transgender patients. Further investigations into this area are needed. In addition, counseling regarding fertility preservation options should become a significant part of the provider-patient conversation before transitioning therapies.

Dual trigger for final oocyte maturation in expected normal responders with a high immature oocyte rate: a randomized controlled trial.

To evaluate whether dual trigger could improve reproductive outcomes in women with low oocyte maturation rates compare to human chorionic gonadotropin (hCG) trigger. This study included expected normal ovarian responders younger than 40 years old whose immature oocyte rate in the previous cycle was more than 50% at the reproductive center from July 2021 to November 2022. A total of 73 patients were enrolled at trigger, including 34 in the hCG trigger group and 39 in the dual trigger group (co-administration of gonadotrophin releasing hormone (GnRH) agonist and hCG, 40 and 34 h prior to oocyte retrieval, respectively). The primary outcome was oocyte maturation rate. There was no significant difference in the number of oocytes retrieved between the two study groups, but the oocyte maturation rate was higher in dual trigger group (84.0% [14.0%] vs. 55.5% [19.8%], p < 0.001). Moreover, there were also higher cumulative pregnancy rate (69.4% vs. 40.0%, p = 0.035) and cumulative live birth rate (66.7% vs. 36.0%, p = 0.022) in dual trigger group. For normal responders with low oocyte maturation rates, the dual trigger may be more effective than the conventional hCG trigger. ClinicalTrials.gov, identifier ChiCTR2100049292.

Factors associated with the collection of isolated immature oocytes during ovarian tissue cryopreservation.

To identify patient characteristics associated with successful isolated immature oocyte retrieval (IsO) during ovarian tissue cryopreservation (OTC) and to determine whether they are predictive of the collection of larger numbers of oocytes. We retrospectively analyzed all patients undergoing OTC with IsO for fertility preservation over three years of activity at a university hospital. Univariate and multivariate analyses were used to identify the patients with the highest and lowest chances of oocyte recovery, and those with the largest numbers of oocytes. We also analyzed the correlation of IsO with the number of ovarian fragments collected and histological parameters. We analyzed 257 consecutive patients undergoing these procedures, at a median age of 17.1 years [0.3-38.3 years]. Isolated oocytes were obtained from 47.1% of patients, and IsO was more likely in patients with ovulatory cycles (63.0% vs. 38.6%; P≤ .001), without chemotherapy before OTC (61.4% vs. 33.1; P< .001) and with non-malignant diseases other than Turner syndrome (77.5%). Oocyte collection failure rates were highest in patients with Turner syndrome (OR 25.0, 95% CI 3.99-157.0; P< .001) or undergoing chemotherapy with alkylating agents before OTC (OR 37.6, 95% CI 8.36-168.8; P< .001). Prepubescent status (P= .043) and large numbers of ovarian fragments (P< .001) were associated with the retrieval of larger numbers of oocytes. Oocyte recovery was correlated with the presence of follicles in the medulla, but not with follicular density. The chances of IsO differ between patients. Identifying patients with the highest chances of success facilitates appropriate resource allocation.

Rapamycin treatment during prolonged in vitro maturation enhances the developmental competence of immature porcine oocytes.

Porcine oocytes undergo in vitro maturation (IVM) for 42-44 h. During this period, most oocytes proceed to metaphase and then to pro-metaphase if the nucleus has sufficiently matured. Forty-four hours is sufficient for oocyte nuclear maturation but not for full maturation of the oocyte cytoplasm. This study investigated the influences of extension of the IVM duration with rapamycin treatment on molecular maturation factors. The phospho-p44/42 mitogen-activated protein kinase (MAPK) level was enhanced in comparison with the total p44/42 MAPK level after 52 h of IVM. Oocytes were treated with and without 10 μM rapamycin (10 R and 0 R, respectively) and examined after 52 h of IVM, whereas control oocytes were examined after 44 h of IVM. Phospho-p44/42 MAPK activity was upregulated the 10 R and 0 R oocytes than in control oocytes. The expression levels of maternal genes were highest in 10 R oocytes and were higher in 0 R oocytes than in control oocytes. Reactive oxygen species (ROS) activity was dramatically increased in 0 R oocytes but was similar in 10 R and control oocytes. The 10 R group exhibited an increased embryo development rate, a higher total cell number per blastocyst, and decreased DNA fragmentation. The mRNA level of development-related (POU5F1 and NANOG) mRNA, oocyte-apoptotic (BCL2L1) genes were highest in 10 R blastocysts. These results suggest that prolonged IVM duration with rapamycin treatment represses ROS production and increases expression of molecular maturation factors. Therefore, this is a good strategy to enhance the developmental capacity in porcine oocytes.

Use of confocal microscopy and intracytoplasmic sperm injection (ICSI) to assess viability of equine oocytes from young and old mares after vitrification.

The impact of vitrification on oocyte developmental competence as a function of donor age remains an important issue in assisted reproductive technologies (ARTs). Equine germinal vesicle (GV) or metaphase II (M(II) oocytes were vitrified using the Cryotop® method. Spindle organization and chromosome alignment were evaluated from confocal imaging data sets of in vivo (IVO) or in vitro (IVM) matured oocytes subjected to vitrification or not. Intracytoplasmic sperm injection (ICSI) from the same groups was used to assess developmental potential. An increase in chromosome misalignment was observed in spindles from older mares when compared to those of younger mares (P < 0.05). When MII oocytes subjected to vitrification were examined following warming, there was no difference in the percentage of oocytes displaying chromosome misalignment. Next, GV oocytes, collected from the ovaries of younger and older mares, were compared between fresh IVM and IVM following vitrification and warming. For nonvitrified samples, an age difference was again noted for spindle organization and chromosome alignment, with a higher (P < 0.05) percentage of normal bipolar meiotic spindles with aligned chromosomes observed in nonvitrified oocytes from young versus older mares. Vitrification led to a reduction of spindle length (P < 0.05) for oocytes from old mares, whether vitrified at GV or MII stages, whereas this effect was not observed in oocytes from young mares except those vitrified at GV and subjected to IVM. Oocyte developmental potential after vitrification was evaluated after ICSI of vitrified and warmed MII or GV oocytes from young mares. From 25 MII oocytes, 18 oocytes were injected with sperm, and six blastocysts were produced, which, upon transfer to mares' uteri, resulted in four pregnancies. Immature (GV) oocytes collected from live mares were also vitrified, warmed, and matured in vitro before ICSI. In this group, nonvitrified, control, and vitrified oocytes did not differ (P > 0.05) with respect to the incidence of maturation to MII, cleavage after ICSI, or blastocyst development. These findings demonstrate an effect of maternal age in an equine model at the level of meiotic spindle integrity and chromosome positioning that is influenced by both the meiotic stage at which oocytes are vitrified and whether meiotic maturation occurred in vivo or in vitro.

Recent Advances in InVitro Maturation (IVM): A Systematic Literature Review

In Vitro Maturation (IVM) is one of the Assisted Reproductive Technologies (ART) that involves the retrieval of immature oocytes from antral follicles in the ovaries, either in a non-stimulated or minimally stimulated state. IVM has drawn the attention of fertility specialists due to its reliability, cost-effectiveness, low risk of Ovarian Hyperstimulation Syndrome (OHSS), and acceptable clinical pregnancy rates Objective: Understanding the Process and Factors Influencing In Vitro Maturation of Human Oocytes (IVM) Method: The research methodology employed in this study is the Systematic Literature Review (SLR) method. Data was collected by documenting all articles based on predetermined inclusion and exclusion criteria. A total of 10 national and international journal articles were utilized for this study, obtained from databases such as Google Scholar, PubMed, and Science Direct, using specific keywords including "In Vitro Maturation" and "human oocyte maturation". The literature search and study selection process followed the PRISMA (Preferred Reporting Items for Systematic Reviews and Meta-Analyses) guidelines. Results: There were no instances of moderate-severe OHSS (ovarian hyperstimulation syndrome) in the IVM group. The fertilization rate in the group receiving growth hormone (GH) was 12.8% higher compared to the control group. Additionally, the blastocyst rate in the GH group was 9.5% higher than that of the control group. Treatment with a concentration of 50 µmol/L ALA (alpha-lipoic acid) significantly accelerated oocyte maturation. It resulted in a significantly higher mitochondrial DNA (mtDNA) copy number in mature oocytes compared to the control group (0 µmol/L ALA). The highest maturation rates were observed in human umbilical cord mesenchymal cells (hUCM) in the in vitro maturation (fIVM) of germinal vesicle (GV) oocytes, while the lowest maturation rates were observed using alpha-Minimum Essential Medium (α-MEM) in the in vitro maturation (vIVM) of GV oocytes. When AMH (anti-Müllerian hormone) was added to the IVM medium, a maturation rate of 100% was achieved in mature oocytes. The inclusion of 50 mmol/L CoQ10 in IVM media was found to reduce the level of oocyte aneuploidy by nearly 50%. The formation rate of metaphase II (MII) stage oocytes was higher in culture medium containing 1.0 mm resveratrol compared to the control group. Subsequently, a total of eight cleavage-stage embryos were frozen after intracytoplasmic sperm injection (ICSI). One year later, when the patient returned for the transfer of cryopreserved embryos, two embryos were thawed and transferred into the uterus. The patient successfully became pregnant, and the twin pregnancy progressed without complications, delivering two healthy full-term baby boys. Conclusion: The occurrence of OHSS was not observed in any of the IVM intervention groups. Supplementation with growth hormone (GH), alpha-lipoic acid (ALA), umbilical cord-derived mesenchymal stem cells (CM-MSC), anti-Müllerian hormone (AMH), and resveratrol in the culture media each demonstrated a significant improvement in human oocyte maturation. Furthermore, resveratrol and coenzyme Q10 were effective in addressing abnormal spindle morphology, irregular chromosome arrangement, and high postmeiotic aneuploidy. It is worth noting that IVM is not exclusively limited to patients with polycystic ovary syndrome (PCOS), as it can yield satisfactory outcomes in patients with autoimmune premature ovarian insufficiency (POI).

The ultrastructural nature of human oocytes’ cytoplasmic abnormalities and the role of cytoskeleton dysfunction

To investigate the structural bases of human oocytes' cytoplasmic abnormalities and the causative mechanism of their emergence. Knowledge of an abnormal oocyte's intracellular organization is vital to establishing reliable criteria for clinical evaluation of oocyte morphology. Laboratory-based study on experimental material provided by a private assisted reproduction clinic. University laboratory and imaging center. A total of 105 women undergoing hormonal stimulation for invitro fertilization (IVF) donated their spare oocytes for this study. Transmission electron microscopy (TEM) was used to analyze the fine morphology of 22 dysmorphic IVF oocytes exhibiting different types of cytoplasmic irregularities, namely, refractile bodies; centrally located cytoplasmic granularity (CLCG); smooth endoplasmic reticulum (SER) disc; and vacuoles. A total of 133 immature oocytes were exposed to cytoskeleton-targeting compounds or matured in control conditions, and their morphology was examined using fluorescent and electron microscopy. The ultrastructural morphology of dysmorphic oocytes was analyzed. Drug-treated oocytes had their maturation efficiency, chromosome-microtubule configurations, and fine intracellular morphology examined. TEM revealed ultrastructural characteristics of common oocyte aberrations and indicated that excessive organelle clustering was the underlying cause of 2 of the studied morphotypes. Inhibition experiments showed that disruption of actin, not microtubules, allows for inordinate aggregation of subcellular structures, resembling the ultrastructural pattern seen in morphologically abnormal oocytes retrieved in IVF cycles. These results imply that actin serves as a regulator of organelle distribution during human oocyte maturation. The ultrastructural analogy between dysmorphic oocytes and oocytes, in which actin network integrity was perturbed, suggests that dysfunction of the actin cytoskeleton might be implicated in generating common cytoplasmic aberrations. Knowledge of human oocytes' inner workings and the origin of morphological abnormalities is a step forward to a more objective oocyte quality assessment in IVF practice.

Oocyte cryopreservation with in vitro maturation for fertility preservation in girls at risk for ovarian insufficiency.

To assess the feasibility and outcomes of oocyte cryopreservation with in vitro maturation (IVM) in post-pubertal girls undergoing fertility preservation (FP) for primary ovarian insufficiency (POI) risk. Ovarian stimulation was performed with an antagonist protocol or progesterone priming. Ultrasound monitoring was performed transabdominally. Oocytes were retrieved transvaginally under IV sedation. Immature oocytes were subjected to IVM for up to 36h. All MII oocytes were vitrified. The main outcome measure was the total number of mature oocytes cryopreserved. The secondary outcome was the increase in the mature oocyte yield after IVM. Indications for FP included mosaic Turner syndrome (mTS; n = 10), malignancy (n = 3), and POI risk (n = 2). The mean ± SD age, antral follicle count (AFC), and AMH levels were 14.2 ± 1.4years, 8 ± 5.2 and 1.3 ± 1.3ng/mL. In girls with mTS, the ovarian reserve was low for age (AFC 7.4 ± 4.7 and AMH 1.4 ± 1.6ng/mL). Oocyte cryopreservation was possible in all girls with a range of 1-27 mature oocytes obtained, even in those who were previously exposed to chemotherapy or with low ovarian reserve, and no surgical complications were encountered. After IVM, the median mature oocyte yield increased significantly from 7.5 to 10.5 (p = 0.001). Oocyte cryopreservation appears to be feasible and safe in girls as young as 12years of age at risk for POI The utility of IVM increases the yield of cryopreserved mature oocytes. Prior exposure to chemotherapy or low ovarian reserve should not be an automatic reason to exclude these girls from FP consideration.

The effects of pretreatment with Cyclosporin A and Docetaxel before vitrification of porcine immature oocytes on subsequent embryo development

In the present study, we attempted to improve the developmental competence of vitrified immature porcine oocytes by the preservation of mitochondrial properties using Cyclosporin A (CsA, inhibitor of mitochondrial membrane permeability transition) and Docetaxel (stabilizer of microtubules, hence mitochondrial distribution). In Experiment 1, Mitotracker red staining revealed reduced mitochondrial activity (MA) in vitrified/warmed oocytes at 0 and 22 h of in vitro maturation (IVM) compared with fresh ones. However, by at 46 h of IVM, MA levels in vitrified oocytes were similar to those in fresh control. Treatment of oocytes with CsA or Docetaxel improved MA at 0 h and 22 h of IVM compared with non-treated vitrified oocytes. However, there were no significant differences among groups in percentages of survival, maturation and embryo development after subsequent IVM and parthenogenetic activation. Nevertheless, a pretreatment with a combination of 10 µg/mL CsA and 0.05 µM Docetaxel improved the blastocyst formation of vitrified oocytes compared with non-treatment counterparts (11.2 ± 1.6% vs 5.9 ± 1.6%, P < 0.05). In conclusion, vitrification reduced mitochondrial activity in GV-stage oocytes during 0–22 h of IVM; however, it was normalized by 46 h IVM. Docetaxel or CsA pretreatment alone did not improve development competence of vitrified oocytes. However, pretreatment with a combination of CsA and Docetaxel could improve blastocyst formation rates.

The Effect of Wharton's Jelly-Derived Conditioned Medium on the In Vitro Maturation of Immature Oocytes, Embryo Development, and Genes Expression Involved in Apoptosis.

Oocyte cytoplasmic maturation is a crucial process during in vitro maturation (IVM), and finding an appropriate IVM medium that promotes oocyte competence is very critical in assisted reproductive technology (ART). The aim of this study was to investigate the effects of umbilical cord Wharton's jelly (WJ-MSCs)-derived conditioned media on the maturation of immature oocytes and their developmental potential in humans after IVM, as well as apoptotic gene expression. A total of 392 germinal vesicle (GV) oocytes were collected from 207 women aged 25-35 years and divided into two IVM groups: (1) control group, which was cultured in CleavTM medium, and (2) experimental group, which was cultured in supernatants of umbilical cord Wharton's jelly as a conditioned medium (CM). First, WJ-MSCs were isolated, and their purity was analyzed. The immunophenotypes of WJ-MSCs were analyzed by flow cytometry. The quantitative expression of BCL2, BAX, and BAG1 in matured oocytes and embryos was evaluated through quantitative real-time polymerase chain reaction (qRT-PCR). Our findings showed that WJ-MSCs have a high proliferating capacity. The purity of the isolated cells was further validated by immunophenotyping, which revealed that their surface antigen expression had phenotypical properties similar to WJ-MSCs. When compared to CD34 and CD45 surface markers, the enlarged cells were positive for CD90, CD105, and CD44. There were significant differences in cytoplasmic maturation of oocytes and embryo quality between the two groups. The mRNA expression levels of BCL-2, BAG1, and BAX in matured oocytes and embryos were also significantly different between the two groups. Therefore, WJ-MSCs medium indicated potential efficacy in terms of ameliorating oocyte maturation and in promoting the development and genes expression of BAX, BCL-2, and BAG1.

Clinical and laboratory factors associated with the presence of dysmorphic oocytes in intracytoplasmic sperm injection cycles.

This study investigated the clinical and laboratory factors associated with the presence of dysmorphic oocytes in intracytoplasmic sperm injection (ICSI) cycles. The study involved 200 ICSI cycles, performed from 2020 to 2021, that yielded at least one mature oocyte. Clinical characteristics and ovarian stimulation methods were compared between 68 cycles with at least one dysmorphic oocyte (the dysmorphic group) and 132 cycles with normal-form oocytes only (the non-dysmorphic group). Dysmorphic oocytes were characterized by dark cytoplasm, cytoplasmic granularity, cytoplasmic vacuoles, refractile bodies in the cytoplasm, smooth endoplasmic reticulum in the cytoplasm, an oval shape, an abnormal zona pellucida, a large perivitelline space, debris in the perivitelline space, or an abnormal polar body. The ages of the women, indications for in vitro fertilization, serum anti-Müllerian hormone levels, and rates of current ovarian endometrioma were similar between the dysmorphic and non-dysmorphic groups. In both groups, the three ovarian stimulation regimens, two types of pituitary suppression, and total gonadotropin dose were employed similarly. However, the dual-trigger method was used more frequently in the dysmorphic group (67.6% vs. 50%, p=0.024). The dysmorphic group contained significantly more immature oocytes and exhibited significantly lower oocyte maturity (50% vs. 66.7%, p=0.001) than the non-dysmorphic cycles. Within the dysmorphic group, significantly lower oocyte maturity was found in the cycles using a dual-trigger, but not in those with a human chorionic gonadotropin trigger. ICSI cycles with dysmorphic oocytes are closely associated with reduced oocyte maturity. This association was observed exclusively in dual-trigger cycles.