MII Oocytes Research Articles

Predictive modeling of oocyte maternal mRNA features for five mammalian species reveals potential shared and species-restricted regulators during maturation.

Oocyte maturation is accompanied by changes in abundances of thousands of mRNAs, many degraded and many preferentially stabilized. mRNA stability can be regulated by diverse features including GC content, codon bias, and motifs within the 3'-untranslated region (UTR) interacting with RNA binding proteins (RBPs) and miRNAs. Many studies have identified factors participating in mRNA splicing, bulk mRNA storage, and translational recruitment in mammalian oocytes, but the roles of potentially hundreds of expressed factors, how they regulate cohorts of thousands of mRNAs, and to what extent their functions are conserved across species has not been determined. We performed an extensive in silico cross-species analysis of features associated with mRNAs of different stability classes during oocyte maturation (stable, moderately degraded, and highly degraded) for five mammalian species. Using publicly available RNA sequencing data for germinal vesicle (GV) and MII oocyte transcriptomes, we determined that 3'-UTR length and synonymous codon usage are positively associated with stability, while greater GC content is negatively associated with stability. By applying machine learning and feature selection strategies, we identified RBPs and miRNAs that are predictive of mRNA stability, including some across multiple species and others more species-restricted. The results provide new insight into the mechanisms regulating maternal mRNA stabilization or degradation.NEW & NOTEWORTHY Conservation across species of mRNA features regulating maternal mRNA stability during mammalian oocyte maturation was analyzed. 3'-Untranslated region length and synonymous codon usage are positively associated with stability, while GC content is negatively associated. Just three RNA binding protein motifs were predicted to regulate mRNA stability across all five species examined, but associated pathways and functions are shared, indicating oocytes of different species arrive at comparable physiological destinations via different routes.

The usefulness of peri-trigger female reproductive hormones (delta-FRH) in predicting oocyte maturation in normal ovarian reserve patients who received in vitro fertilization-embryo transfer: a retrospective study.

To evaluate the efficacy of peri-trigger female reproductive hormones (FRHs) in the prediction of oocyte maturation in normal ovarian reserve patients during the in vitro fertilization-embryo transfer (IVF-ET) procedure. A hospital database was used to extract data on IVF-ET cases from January 2020 to September 2021. The levels of female reproductive hormones, including estradiol (E2), luteinizing hormone (LH), progesterone (P), and follicle-stimulating hormone (FSH), were initially evaluated at baseline, the day of the trigger, the day after the trigger, and the day of oocyte retrieval. The relative change in E2, LH, P, FSH between time point 1 (the day of trigger and baseline) and time point 2 (the day after the trigger and day on the trigger) was defined as E2_RoV1/2, LH_RoV1/2, P_RoV1/2, and FSH_RoV1/2, respectively. Univariable and multivariable regression were performed to screen the peri-trigger FRHs for the prediction of oocyte maturation. A total of 118 patients were enrolled in our study. Univariable analysis revealed significant associations between E2_RoV1 and the rate of MII oocytes in the GnRH-agonist protocol group (p<0.05), but not in the GnRH-antagonist protocol group. Conversely, P_RoV2 emerged as a potential predictor for the rate of MII oocytes in both protocol groups (p<0.05). Multivariable analysis confirmed the significance of P_RoV2 in predicting oocyte maturation rate in both groups (p<0.05), while the association of E2_RoV1 was not significant in either group. However, within the subgroup of high P_RoV2 in the GnRH-agonist protocol group, association was not observed to be significant. The C-index was 0.83 (95% CI [0.73-0.92]) for the GnRH-agonist protocol group and 0.77 (95% CI [0.63-0.90]) for the GnRH-antagonist protocol group. The ROC curve analysis further supported the satisfactory performance of the models, with area under the curve (AUC) values of 0.79 for the GnRH-agonist protocol group and 0.81 for the GnRH-antagonist protocol group. P_RoV2 showed significant predictive value for oocyte maturation in both GnRH-agonist and GnRH-antagonist protocol groups, which enhances the understanding of evaluating oocyte maturation and inform individualized treatment protocols in controlled ovarian hyperstimulation during IVF-ET for normal ovarian reserve patients.

Ovarian response determines the luteinizing hormone suppression threshold for patients following the gonadotrophin releasing hormone antagonist protocol: A retrospective cohort study

BackgroundOvarian reactivity to gonadotrophin stimulation varies, and individual adjustments to the timing and dose of gonadotrophin-releasing hormone (GnRH) antagonist administration are necessary to prevent excessive increases and decreases in luteinizing hormone (LH) levels in patients with different ovarian response following the GnRH antagonist (GnRH-A) protocol. The present study aims to investigate optimal LH suppression thresholds for patients with normal ovarian response (NOR), high ovarian response (HOR), and poor ovarian response (POR) following the GnRH-A protocol respectively. MethodsA total of 865 in vitro fertilization (IVF) cycles using a flexible or fixed GnRH-A protocol were included. Patients were categorized into the HOR, NOR, or POR group according to their anti-Müllerian hormone (AMH) levels. Then, patients in each group were stratified into one of four subgroups according to the quartile (Q1-Q4) of the basal LH level to LH on triggering day ratio (bLH/hLH). The primary outcomes were the clinical pregnancy and live birth rates, and the secondary outcomes were the number of oocytes retrieved, MII oocytes, two pronucleus (2PN) embryos, and good-quality embryos. ResultsThere were 526 patients with NOR, 180 with HOR, and 159 with POR. Basal LH level, LH on triggering day and bLH/hLH were identified as independent predictors of clinical pregnancy rate and live birth rate by logistics regression analysis. Compared to those with NOR, patients with POR had the lowest embryo implantation rate (22.6% vs. 32.8%, P < 0.05), clinical pregnancy rate (32.3% vs. 47.3%, P < 0.05) and live birth rate (22.6 vs. 37.8%, P < 0.05) of fresh embryo transfer (ET). The embryo implantation, clinical pregnancy and live birth rates of frozen embryo transfer (FET) were not significantly different among the three groups. In the subgroup analysis, patients with HOR had the highest embryo implantation rate (51.6%, P < 0.05), clinical pregnancy rate (68.4%, P < 0.05) and live birth rate (52.6%, P < 0.05) of ET in Q3, with a bLH/hLH ratio of 2.40–3.69. In the NOR group, the embryo implantation rate (41.9%, P < 0.05), clinical pregnancy rate (61.5%, P < 0.05) and live birth rate (50.8%, P < 0.05) of ET and live birth rate (53.1%, P < 0.05) of FET were highest in Q2, with a bLH/hLH ratio of 1.29–2.05. Patients with POR had the highest clinical pregnancy rate (57.1%, P < 0.05) and live birth rate (42.9%, P < 0.05) of ET in Q2, with a bLH/hLH ratio of 0.86–1.35. ConclusionsIn the present study, the bLH/hLH ratio represented the LH suppression threshold. The subgroup analysis of HOR, NOR and POR showed that, the LH suppression threshold varies according to ovarian response. We recommend LH suppression thresholds of 2.40–3.69 for HOR, 1.29–2.05 for NOR, and 0.86–1.35 for POR to obtain the highest clinical pregnancy rate and live birth rate. This study provides comprehensive and precise references for clinicians to monitor LH levels individually during controlled ovarian stimulation (COS) according to the patient’s ovarian response following the GnRH-A protocol.

Correlation between controlled ovarian stimulation protocols and euploid blastocyst rate in pre-implantation genetic testing for aneuploidy cycles

BackgroundSeveral studies have explored which COS protocol yields a higher blastocyst euploidy rate, but findings have been inconsistent. The present study aimed to explore whether controlled ovarian stimulation (COS) protocols was associated with euploid blastocyst rate in pre-implantation genetic testing for aneuploidy (PGT-A) cycles.MethodsThe study was a retrospective study where data were obtained from three reproductive medicine centers. The study included PGT-A cycles with the GnRH-a, GnRH-ant, or PPOS protocols, and the data on patient demographics, protocols, and embryonic outcomes were collected for the PGT-A cycles performed between January 2019 and August 2022.ResultsThis study included 457 PGT-A cycles from three reproductive medicine centers, with 152, 126, and 179 cycles performed using the PPOS, GnRH-a, and GnRH-ant protocols, respectively. The baseline characteristics of the three groups show no significant differences were observed in female BMI, infertility type, and infertility duration among the PPOS, GnRH-a, and GnRH-ant protocol groups. The study found no significant association between Gn dosage, Gn duration, and blastocyst euploidy. The mean number of euploidy blastocysts in PPOS protocol was significantly lower than that of GnRH-a protocol and GnRH-ant protocol (0.75 ± 0.92 vs. 1.79 ± 1.78 vs. 1.80 ± 1.67). The euploidy rate per biopsy blastocyst (48.4% vs. 49.1% vs. 33.1%), per oocyte retrieved (15.0% vs. 14.7% vs. 10.5%), and per MII oocyte (17.7% vs. 16.4% vs. 11.7%) were significantly higher in the cycles using the GnRH-ant and GnRH-a protocols than that of PPOS protocol group. Regression analyses indicated that, compared with the PPOS protocol, the GnRH-ant protocol was positively associated with the euploid blastocyst rate and the mean number of euploid blastocysts, whereas the GnRH-a protocol showed no such relationship.Limitations and reasons for cautionThe main limitation of this study was the retrospective design. Although this study also used other tests to account for confounding factors and reduce potential bias, multiple tests have its own weaknesses.ConclusionsGnRH-ant protocol was the most effective for PGT-A cycles. The findings emphasize the need for personalized treatment strategies, considering patient demographics, and optimizing COS protocols to enhance the chances of successful outcomes in ART procedures.

#239 : Granulocyte-Macrophage-Colony Stimulating Factor Improve Embryo Development, Mitochondrial Function and Spindle Alignment in a Mouse Model of Advanced Maternal Age

Background and Aims: We have previously shown that GM-CSF can improve embryo development when added during in vitro oocyte maturation (IVM) as well as increase pregnancy and birth rates following embryo transfer in mice. The aim of the present study was to determine whether GM-CSF during IVM can improve embryo development in a mouse model of advanced maternal age (AMA) Method: 12–14-month-old CBAF1 female mice were injected with 5IU of PMSG and cumulus-oocyte complexes (COCs) aspirated 46–48 h post-injection. Ten COCs were cultured per 50 [Formula: see text]L drop in bicarbonate-buffered [Formula: see text]-MEM containing 3 mg/ml BSA, 1 mg/ml Fetuin and 5 mIU/mL FSH plus 0 or 10 ng/ml of GM-CSF. Zygotes were cultured in Cleave Media and cleavage and blastocyst rates were determined. Blastocysts were differentially stained on Day 5 to determine cell numbers. To determine mitochondrial membrane potential (MMP) COCs were incubated for 15 minutes in HEPES buffered alpha-MEM media with JC-1 dye for 15 minutes. To examine spindle formation and chromosomal alignment, MII oocytes were fixed and then incubated with FITC conjugated mouse anti-[Formula: see text]-tubulin for 40 minutes at 25°C and chromosomes stained using Hoechst. Fluorescence was visualised using Cell Voyager CV1000 Confocal Scanner and Z-stack images were analysed using Fiji Image J software. Data were analysed using a univariate general liner model in IBM SPSS. Results: GM-CSF during IVM increased Day 5 blastocyst rate by 21% (P&lt;0.01) and the proportion of expanded blastocysts by 75% (P&lt;0.05). Blastocyst inner cell mass was increased by 23% (P&lt;0.05) and total cell number by 14% (P&lt;0.05). MMP was increased across all fields by three-fold (P&lt;0.05). GM-CSF did not affect spindle formation but tended to reduce chromosomal misalignment by 17% (P&gt;0.05). Conclusion: We have shown that the addition of GM-CSF during IVM can improve embryo development and mitochondrial activity in a mouse of AMA.

Non-inferiority of progestin-primed ovarian stimulation versus GnRH antagonist protocol: A propensity score-weighted analysis

PurposeTo evaluate the effectiveness of the progestin-primed ovarian stimulation (PPOS) protocol versus the gonadotropin-releasing hormone antagonist (GnRH-ant) protocol in ovarian stimulation. MethodsIn this retrospective cohort study, we included 804 patients who were treated between January 1st, 2022, and July 1st, 2023. Outcomes of ovarian stimulation were compared between the PPOS (n = 206) and GnRH-ant (n = 598). The primary outcome was the number of good cleavage embryos. ResultsBaseline characteristics were comparable in both groups. In both unadjusted and adjusted analysis, the mean number of good cleavage embryos in PPOS (6.33) was non-inferior to GnRH-ant (6.44; unadjusted ratio of two means 1.02, 95%CI 0.92, 1.13). The trigger-day estradiol level in patients with PPOS was higher than in patients with GnRH-ant (4,420 vs 3,830 pg/ml, respectively) despite similar total follicle stimulating hormone dose and fewer days of ovarian stimulation. The number of oocytes, MII oocytes, cleavage and blastocyst embryos were comparable between the two protocols. After the first transfer of embryos, the clinical pregnancy rate and implantation rate were higher in the PPOS group, while the pregnancy rate and ongoing pregnancy were not significantly different. None of the PPOS patients had an unexpected LH surge, and serum LH levels decreased slightly during ovarian stimulation. ConclusionsThe PPOS protocol with dydrogesterone provided similar embryo outcomes to the GnRH-ant protocol, with notable distinctions in clinical pregnancy and implantation rate. The serum LH concentration during ovarian stimulation using PPOS was well-controlled.

In vitro reduction of bovine oocyte ATP production with oligomycin affects embryo epigenome.

Epigenetic programming is a crucial process during early embryo development that can have a significant impact on the results of assisted reproductive technology (ART) and offspring health. Here we show evidence using a bovine in vitro experiment that embryo epigenetic programing is dependent on oocyte mitochondrial bioenergetic activity during maturation. This study investigated if oocyte and early embryo epigenetic programming are dependent on oocyte mitochondrial ATP production. A bovine in vitro experiment was performed in which oocyte mitochondrial ATP production was reduced using 5 nmol/L oligomycin A (OM; ATP synthase inhibitor) during in vitro maturation (IVM) compared to control (CONT). OM exposure significantly reduced mitochondrial ATP production rate in MII oocytes (34.6% reduction, P = 0.018) and significantly decreased embryo cleavage rate at 48 h post insemination (7.6% reduction, P = 0.031). Compared to CONT, global DNA methylation (5mC) levels were decreased in OM-exposed MII oocytes (9.8% reduction, P = 0.019) while global histone methylation (H3K9me2) was increased (9.4% increase, P = 0.024). In zygotes, OM exposure during IVM increased 5mC (22.3% increase, P < 0.001) and histone acetylation (H3K9ac, 17.3% increase, P = 0.023) levels, while H3K9me2 levels were not affected. In morulae, 5mC levels were increased (10.3% increase, P = 0.041) after OM exposure compared to CONT, while there was no significant difference in H3K9ac and H3K9me2 levels. These epigenetic alterations were not associated with any persistent effects on embryo mitochondrial ATP production rate or mitochondrial membrane potential (assessed at the four-cell stage). Also, epigenetic regulatory genes were not differentially expressed in OM-exposed zygotes or morulae. Finally, apoptotic cell index in blastocysts was increased after OM exposure during oocyte maturation (41.1% increase, P < 0.001). We conclude that oocyte and early embryo epigenetic programming are dependent on mitochondrial ATP production during IVM.

#282 : Maximizing IVF Success Rates: Optimized Simulation Using a Novel Decision-Support Tool

Background and Aims: A mathematical procedure and a decision support tool, Opt-IVF, were developed to overcome the shortcomings of the current IVF practice of dosage profile determination by trial and error, which can provide a personalized model and optimized drug treatment. This paper describes the decision support tool for improving outcomes of IVF treatment for both antagonist and agonist protocols used in real practice. We present the results of two clinical trials. Method: Our earlier work developed models for agonist and antagonist protocols based on an analogy between superovulation and batch crystallization. This model was customized for each patient using two days of follicular data, and the optimal control method was applied to optimize the dosage profile. Validation of the model was conducted with retrospective data from 150 patients from India and 50 from the United States. Opt-IVF reduced total hormone dosage by 20 to 60%, reduced US testing by 50 to 75%, and provided a higher number of oocytes and high-quality embryos. Results: The primary outcome measures studied for two clinical trials with Opt-IVF include total hormonal dosage, % follicles retrieved, MII oocytes, Embryos, and Grade A Embryos. Results showed that Opt-IVF reduces drug dosage and testing and increases the number of embryos, Grade A embryos, and high-quality blastocysts significantly. Subgroup analysis showed improvements in outcomes were more significant for both Poor-responder and PCOS patients than for normal responders. Pregnancy rates were significantly higher with Opt-IVF patients than with standard protocol control patients. Conclusions: A mathematics-based approach to dosage profile determination results in optimal mature follicles, better embryos, and higher pregnancy with reduced dosage and the need for testing.

#50 : Effect of Dual vs. Single (hCG) Trigger in Maturation Rate of Oocytes in ART

Background and Aims: Due to its similar molecular structures and biological activities, human chorionic gonadotropin (hCG) is used as a substitute for luteinizing hormone (LH) in ART. The weaknesses of using hCG as a trigger are the possibility of ovarian hyperstimulation syndrome in high responders and lacked FSH activity which promotes LH receptor formation in luteinizing granulosa cells, nuclear maturation, and cumulus expansion. GnRH agonist (GnRHa) reduces the development of OHSS and induces a physiological surge similar to the natural menstrual cycle. Because of shorter amplitude and duration of GnRHa-induced LH surge, GnRHa-alone trigger results in a higher early miscarriage rate. Combining these two agents; dual trigger increases oocyte maturation with reduced OHSS. Here we present reports in our clinic to differentiate the dual vs. hCG trigger results. Method: We retrospectively analyzed the data from 68 patients who had undergone antagonist protocol from January 2021-January 2023. The final oocyte maturation was undertaken with a dual trigger with GnRHa (Buserelin 0.3 mg) combined with hCG (r-hCG 250 [Formula: see text]g) ([Formula: see text] = 33) or hCG alone ([Formula: see text] = 35). Results: The mean age of both groups was 34 years old. The AMH level of the dual group ranged from 0.06 – 7.17 ng/mL, and in hCG alone group ranged from 0.16 – 12.3 ng/mL. Women in both groups got similar amounts of follicles and MII oocytes; dual group (14.8 and 12.72 vs. 14.9 and 13). The retrieval rate of oocytes was identical in both groups (86.6% vs. 89.9%). However, the maturation rate of dual group was slightly lower than the hCG group (77% vs. 82%), but it was statistically insignificant (independent t-test p 0.99). Conclusion: We showed that dual trigger was not superior to hCG-alone trigger in terms of maturation rate of oocytes. Analysis of larger data is needed to show the real advantages.

#72 : Embryo Quality and Pregnancy Outcome from Oocytes with Smooth Endoplasmic Reticulum (SER) Aggregates

Background and Aims: Calcium stored and released by the Smooth Endoplasmic Reticulum - SER which has an important role in oocyte maturation, fertilization, and embryo development. The presence of SER aggregate result in the disturbance of calcium stores and its occillation which in turn affect fertilization, embryo develompent, and increasing risk of abnormal cromoseme outcome. Alpha-ESHRE Consensus (2011), strongly recomended to not perform insemination on any oocytes with SER. In the other hand, some previous studies show that healthy babies had been born from oocytes presenting this dymorphism. The aim of this study was to evaluate embryo quality and pregnancy outcome patients derived from oocytes with SER. Method: This study was a crosssectional analysis from 2017-2019 of 223 MII oocytes with SER from 82 women underwent Intra cytoplasmic sperm injection - ICSI. Prior to ICSI, each oocyte was evaluated for the presence of SER Aggregates. The embryological outcome measures were the fertilization rate, embryo quality day 3 and day 5, and pregnancy rates. Results: This study result shown that from 223 oocytes with SER, 89.5% oocytes can be fertilized. Day 3 embryo 66.6% ± 0.000 qualified as good embryo, but only 29.1% ± 0.000 on day 5 embryo. The Pregnancy rate was 30.0%, with 2 miscarriages. It is important to give a well information to the patients on the risk of birth malformations as well as whether a decreased chance of pregnancy exists. Conclusion: Oocyte with SER still has the ability to develop and has the potential to support pregnancy.

#345 : Progestin-Primed Ovarian Stimulation Protocol Slightly Improves the IVF Funnel in Patients with Poor Ovarian Response

Background and Aims: Controlled Ovarian Stimulation (COS) is a crucial first step in the In Vitro Fertilization (IVF) or Intra-Cytoplasmic Sperm Injection (ICSI) process. Progestin-primed ovarian stimulation (PPOS) has been recently introduced as an alternative to traditional COS regimens. However, the efficacy of PPOS regimen among infertile women with poor ovarian response (POR) remains unclear. This study aims to evaluate the effect of PPOS regimen on the chance of success throughout the IVF process in Vietnamese infertile women with POR. Method: This randomized controlled trial included 120 infertile women with POR (according to POSEIDON criteria) who received IVF at Tam Anh Fertility Center (Hanoi, Vietnam). PPOS and GnRH_ant regimens were randomly applied to the participants (n=60 for each group). A chain of 4 successive Binomial regression models has been applied to estimate the chance of success for 4 steps: oocyte maturation, fertilization and achieving good quality day-3 and day-5 embryos. The effect of PPOS in comparison with GnRH_ant protocol was evaluated by marginal risk-ratio (RR) from each model. Results: Chances of successfully achieving the MII oocytes, fertilization, high-quality D3 and D5 embryos were all moderately higher when using the PPOS regimen, compared with those of GnRH_ant protocol (95% confidence intervals of RR were respectively 0.88–1.07, 0.89–1.14, 0.96–1.14 and 0.74–1.73). Conclusion: Although not yet reached a statistical significance, those results indicated that using PPOS protocol may slightly improve the chance of success in 4 first critical steps of the IVF process for infertile women with POR.

#230 : The Effectiveness of Spermatid Injection (ROSI, ELSI) Versus Spermatozoa Injection (ICSI) in Infertility Treatment for Men with Non-Obstructive Azoospermia (NOA)

Background and Aims: About 1% of men suffer from azoospermia, a condition of having no sperm in the ejaculate. Non-obstructive azoospermia, which affects 60% of azoospermia patients, is the most severe form of male infertility and requires testicular sperm extraction for sperm retrieval. Despite a 50% success rate, if no mature sperm is found, NOA men have no choice become biological fathers. To overcome this, researchers have attempted to use spermatid injection techniques, such as round spermatid injection (ROSI) or elongating/elongated spermatid injection (ELSI), to give hope for pregnancy. Spermatids are immature sperm but contain a haploid genome as spermatozoa. The aim of this prospective cohort study was to evaluate the effectiveness of ROSI and ELSI in cases where spermatozoa were unavailable. Method: Ninety-nine men diagnosed with NOA were recruited for the study. When the male partners lack spermatozoa in the testicular biopsy but have normal good-looking spermatids, they are consulted to join in the research. The intervention group received ROSI and ELSI, while the control group received spermatozoa injection into oocytes (ICSI). The primary outcome measures were fertilization, embryo development, clinical pregnancy, and live birth rate. Results: The overall fertilization rate (2PN) per MII oocyte injected after ROSI was low at 31.0% compared to ELSI and ICSI at 67.7% and 77.2%, respectively. The pregnancy rate after ROSI was also low, with 3.3% per transferred cycle and 2.2% per embryo transferred. The pregnancy rate of ROSI was much lower than ELSI (42.9%) and ICSI (55.6%). Normal pregnancy and live birth resulted in three groups (Table 1). Conclusion: ROSI and ELSI can be considered alternative methods of infertility treatment in cases where spermatozoa are unavailable.

Cut-off point of mature oocyte for routine clinical application of rescue IVM: a retrospective cohort study

BackgroundThe rescue in vitro mature(Rescue IVM) technique allows the use of immature oocytes collected in conventional COH to obtain more mature oocytes for fertilization through in vitro maturation. Some studies have shown that Rescue IVM could improve clinical outcomes in patients undergoing IVF/ICSI, but the effectiveness and the indications for the clinical application of this technique remain controversial. It remains to be studied whether Rescue IVM should be universally applied in all conventional IVF/ICSI cycles.MethodThis is a large retrospective cohort study that included a total of 22,135 female patients undergoing their first IVF treatment cycles. The effect of the number of mature oocytes(metaphaseII[MII]) on the cumulative live birth rate was investigated in a population with routine IVF/ICSI first. The receiver operating characteristic curve(ROC) analysis was used to explore the cut-off point of the number of MII affecting CLBR. Secondly, Patients undergoing ICSI with Rescue IVM were included in the analysis with those who underwent ICSI only during the same period, grouped according to the MII cut-off values. Multi-factor binary logistic regression and inverse probability weighting (IPW) were used to investigate whether Rescue IVM influenced the final cumulative live birth rate(CLBR).ResultsThe CLBR increased with the number of MIIoocytes (P < 0.001). The ROC analysis showed the cut-off point for the number of MIIoocytes to have a significant effect on CLBR was 9 (sensitivity 0.715, specificity 0.656). Furthermore, 912 patients who underwent ICSI with Rescue IVM were included and compared to those who underwent ICSI only during the same period, and found Rescue IVM significantly increased the number of available MIIoocytes. For patients with MII numbers < 9, Rescue IVM significantly improves their clinical pregnancy rate(55.6% vs. 46.7%, P = 0.001) and CLBR(65.4% vs. 48.1%, P < 0.001), but not for those patients with MII numbers ≥ 9.ConclusionThis study further clarifies the candidates for the application of Rescue IVM technique: patients with an MII oocytes < 9 in a conventional IVF/ICSI cycle. In contrast, it is not necessary for patients who already have sufficient mature oocytes(≥ 9), to avoid over-medication.

Supplementation of media with gamma-oryzanol as a novel antioxidant to overcome redox imbalance during bovine oocyte maturation invitro.

This study evaluated the effect of supplementing IVM media with γ-oryzanol (ORY), a nutraceutical derived from rice bran oil, on the development of bovine oocytes and hindering the compromising effect of redox imbalance. An invitro model of the bovine cumulus-oocyte complex was used for the evaluation of nuclear maturation and development. Antioxidant activity was investigated by assessing the level of ROS (Reactive Oxygen Species) and GSH (glutathione) in oocytes and quantitative changes in gene expression in matured oocytes and their respective cumulus cells. ORY supplementation increased the proportion of MII oocytes, cleaved embryos, and total blastocysts (p < .05) and was linked to higher and lower levels of intracellular GSH and ROS, respectively (p < .05). The treated oocytes and their respective cumulus-granulosa cells showed a modulation in the expression of genes related to apoptosis (downregulation of BAX and CHOP) and oxidative stress (upregulation of NRF2, CAT, and SOD). Also, relative upregulation of OCT-4 and IGF2R in treated oocytes was concomitant with higher subsequent development in terms of cleavage and total blastocyst rates (p < .05). Based on our findings, it appears that ORY supplementation can improve the nuclear maturation and development of bovine oocytes into blastocysts and augment their enzymatic and non-enzymatic antioxidant systems, maintaining the Redox balance and high enzymatic activity against ROS generation.

Mitofusin 1 as Marker of Oocyte Maturation in Relevance to ICSI Outcome in Infertile Females

Pro-fusion proteins as mitofusins-1 are required for controlling mitochondrial shape which determined by a dynamic balance between organelle fusion and fission and also supports oocyte development. When compare with somatic cell, mitochondria of oocyte are tiny and circular in presence. The aim is to study the mitofusin-1 in the follicular fluid as a marker for evaluating of oocyte maturation in women undergoing ICSI cycles. Fifty infertile females were included in cross-section study was undergoing ICSI procedure with age 20 to 42 years. After retrieval of oocyte and the number of oocytes was recorded by embryologist and follicular fluid Samples were used for the measurement of Mitofusin 1. Mitofusin 1 levels by ELISA kit (Mybiosource /USA).The results showed considerable higher mitofusin1levels in patients with good oocytes quality (3.71 ± 1.35 vs. 2.45 ± 1.12 &amp; p=0.001). There were much positive correlations between follicular fluids mitofusin-1 with both total oocytes count (r= 0.374 &amp; p= 0.007) and MII oocytes (r=0.383 &amp; p=0.006); and there was no much correlation between mitofusin-1 levels with MI oocyte (r= - 0.100 &amp; p= 0.490), germinal vesical oocyte (r=0.103 &amp; p= 0.475) and fertilization rate (r= 0.054 &amp; p= 0.711). High follicular fluid levels of Mitofusin 1 may positively impact oocyte development and pregnancy rate.

The effect of letrozole as an adjunct in GnRH-antagonist protocol on IVF/ICSI outcome in women with endometriosis: a randomized clinical trial

BackgroundTo evaluate the effect of adding letrozole to the antagonist ovarian stimulation protocol (COS) on in-vitro fertilization/intracytoplasmic sperm injection (IVF/ICSI) outcome in endometriosis patients.MethodsThis randomized clinical trial was carried out in the department of infertility treatment at Arash Women’s Hospital from May 2019 to May 2021. The eligible women with normal ovarian reserve tests who had endometriosis diagnosis and underwent IVF/ICSI cycles were evaluated. A flexible regimen of GnRH-antagonist protocol was used for COS. In the experimental (n = 34), the patients received 5 mg letrozole daily for the first 5 days in combination with 150 IU of recombinant follicle-stimulating hormone (rFSH). In the control group (n = 30), the patients received only the same dose of rFSH. The treatment cycle was compared between groups.ResultsAnalysis of demographic characteristics, severity of endometriosis, and baseline hormonal tests of patients showed that the two groups were similar and comparable. The means of total used gonadotropins ampoules and serum E2 level on oocyte trigger day in the letrozole group were significantly lower than those of in the control group (P = 0.03 and P = 0.004, respectively). No statistically significant difference in terms of the total number of retrieved and MII oocytes as well as the total numbers of obtained and top-quality embryos, and cryopreserved embryos was found.ConclusionThe co-treatment of letrozole with gonadotropins during the antagonist protocol was associated with a reduction in the total dose of gonadotropins, although it had no effect on the oocyte or embryo yield in patients with endometriosis.Trial registrationThe study was registered in the Iranian Registry of Clinical Trials on 2018 -07-13 (IRCT20150310021420N4 at www.irct.ir, registered while recruiting).

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.

Effect of growth hormone administration on ameliorating pregnancy outcome in women with advanced maternal age and exploration of its optimized utilization.

Age-related fertility decay is a great challenge for clinicians. Growth hormone (GH) supplementation has been studied as an adjuvant since late 1980s. However, it has not come to a consensus on the GH administration due to the ambiguous efficacy among studies with different enrolled population and dosage regime. A self-controlled retrospective study was conducted on women with advanced maternal age who underwent at least a previous cycle without GH (GH-) and a subsequent cycle with GH co-treatment (GH+). The ovarian stimulation parameters and outcomes were compared between the two cycles and logistical analysis was applied to further explore the association between GH administration protocol as well as other clinical parameters and cumulative live birth in GH+cycle. A total of 150 women aged 35-43 were included. The number of oocytes retrieved, MII oocytes, 2PNs, transferrable embryos and good-quality embryos in GH+ significantly increased (p < 0.001). The proportion of cycles with no transferrable embryos was significantly reduced in GH+ cycle compared with previous GH- cycle (3 vs. 32; p < 0.001). GH co-treatment cycles showed significantly higher clinical pregnancy rates (43.75% vs. 6.06%; 38.35% vs. 12.04%, p < 0.001), live birth rates (29.17% vs. 0; 27.07% vs. 0, p < 0.001) in both fresh and frozen-thawed embryo transfer cycle. Cumulative live birth rate of the GH+ cycle reached 33.33%. Use of GH prior to Gn stimulation and lasting until the hCG day seemed to achieve a higher successful live birth rate (OR 2.312, 95%CI 1.074-5.163, p=0.032). GH supplementation could ameliorate pregnancy outcome in women with advanced maternal age. Dosage regimen of long-term pretreatment prior to Gn stimulation (4 IU every other day) and 4 IU per day until hCG day may of greater efficacy compared with concurrent administration with Gn. Additionally, it's worthy of exploring whether an individualized dosage regimen based on the IGF or IGFBP level of patient would be more reasonable and effective. More well-designed prospective trials with large sample size and fundamental experiments on the mechanism are required to testify findings above.

Gonadotropin Versus Gonadotropin/Letrozole Protocol in Previously Failed Antagonist Cycles in Patients with Low Prognosis

Objective: Letrozole is an aromatase inhibitor which has been used for ovulation induction. Our aim is to evaluate the effectiveness of adding letrozole to gonadotropins in the early follicular phase of gonadotropin-releasing hormone antagonist (GnRH-a) protocol in patients with poor ovarian response who had failed in the preceding GnRH-a cycle.&#x0D; Materials and Method: Ninety-eight patients with poor ovarian response who had previously failed GnRH-a cycle were included. Patients (n; 58) who were treated with letrozole plus gonadotropins (LzGA) were compared with patients (n; 40) who received gonadotropins (GT). The number of total oocytes retrieved, the number of MII oocytes and fertilized oocytes, fertilization and implantation ratios, the rate of cycle cancellation as well as clinical pregnancy and live birth rates&#x0D; Results: The LzGA group had a significantly shorter duration of GnRH-a stimulation and higher progesterone level at trigger day (p=0.005, p=0.049 respectively). Although letrozole administration demonstrated lower estradiol levels at trigger day and the total dose of gonadotropins used were lower in LzGA group, this was statistically non-significant (p=0.13, p=0.13 respectively).&#x0D; Conclusion: Adding letrozole to gonadotropins in GnRH-a protocol in patients with poor prognosis did not improve pregnancy outcomes. But it seems to decrease IVF costs by reducing the GnRH-antagonist and gonadotropin dosage.

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.