Fertilization Rate Of Oocytes Research Articles

The effects of decreased ovarian reserve on in vitro fertilization outcome

Objective: In this study, we aimed to compare the effects of microdose and antagonist regimens on IVF-ICSI outcome. 
 Material and Methods: The study was conducted at the XXXX University Medical Faculty Training and Research Hospital IVF Clinic during the period of 2018 and 2022. The files of the patients were reviewed retrospectively and age, duration of infertility, basal FSH level, body mass index, antral follicle number (AFC), antimullerian hormon (AMH) level, retrieved oocyte number, metaphase 2 oocyte number, fertilization and pregnancy rates were evaluated. The patients in Group 1 (n=124) were treated by using gonadotropin releasing hormone (GnRH) microdose protocol and in Group 2 (n=136) were treated by using antagonist protocol. 
 Results: There was no significant difference between the groups in terms of the characteristics of the patients such as age, infertility duration, basal FSH level body mass index, AFC and AMH level. The retrieved oocyte number, metaphase 2 oocyte number, fertilization rates and pregnancy rates were also similar.
 Conclusion: Our results demonstrated that microdose and antagonist regimens have similar effect in poor reponder women on IVF-ICSI outcome.

Human oocyte area is associated with preimplantation embryo usage and early embryo development: the Rotterdam Periconception Cohort

PurposeTo investigate the association between oocyte area and fertilization rate, embryo usage, and preimplantation embryo development in order to establish if oocyte area can be a marker for optimal early embryo development.MethodsFrom 2017 to 2020, 378 couples with an indication for IVF (n = 124) or ICSI (n = 254) were included preconceptionally in the Rotterdam Periconception Cohort. Resulting oocytes (n = 2810) were fertilized and submitted to time-lapse embryo culture. Oocyte area was measured at the moment of fertilization (t0), pronuclear appearance (tPNa), and fading (tPNf). Fertilization rate, embryo usage and quality, and embryo morphokinetics from 2-cell stage to expanded blastocyst stage (t2-tEB) were used as outcome measures in association with oocyte area. Oocytes were termed “used” if they were fertilized and embryo development resulted in transfer or cryopreservation, and otherwise termed “discarded”. Analyses were adjusted for relevant confounders.ResultsOocyte area decreased from t0 to tPNf after IVF and ICSI, and oocytes with larger area shrank faster (β − 12.6 µm2/h, 95%CI − 14.6; − 10.5, p < 0.001). Oocytes that resulted in a used embryo were larger at all time-points and reached tPNf faster than oocytes that fertilized but were discarded (oocyte area at tPNf in used 9864 ± 595 µm2 versus discarded 9679 ± 673 µm2, p < 0.001, tPNf in used 23.6 ± 3.2 h versus discarded 25.6 ± 5.9 h, p < 0.001). Larger oocytes had higher odds of being used (oocyte area at tPNf ORused 1.669, 95%CI 1.336; 2.085, p < 0.001), were associated with faster embryo development up to the morula stage (e.g., t9 β − 0.131 min, 95%CI − 0.237; − 0.025, p = 0.016) and higher ICM quality.ConclusionOocyte area is an informative marker for the preimplantation development of the embryo, as a larger oocyte area is associated with higher quality, faster developing embryos, and higher chance of being used. Identifying determinants associated with oocyte and embryo viability and quality could contribute to improved preconception care and subsequently healthy pregnancies.

The impact of oocyte cryopreservation time in oocyte donation on the clinical success rate

To evaluate the impact of the cryopreservation time of vitrified oocytes on the success rates in oocyte donation cycles. A retrospective study was carried out on 156 cycles with donated oocytes from January 2012 to September 2021. All the cycles were sorted according to the storage time of the oocytes (25 in the group 1: < 3 months, 32 in the group 2: between 3 and 6 months, 39 in the group 3: between 6 and 12 months, 38 in the group 4: between 12 and 24 months and 22 in the group 5: > 24 months). Clinical outcomes after ART, survival rates at thawing and oocyte fertilization rates were compared between the different cohorts stratified according to oocyte storage duration. A binary multivariate logistic regression was performed adjusting for the identified confounders. Prolonged storage time of vitrified oocytes had an effect on their survival post-thawing rates, but no significant effect was identified on fertilization rates or clinical outcomes. After adjusting for the confounders, the relationships between clinical outcomes and oocytes storage time did not reach statistical significance. Our study was characterized by a limited cohort with data from a single ART center. Our study doesn't highlight any significant difference in the use of long-stored vitrified oocytes (more than 2 years) on clinical issues in ART. The conclusion of our study needs to be verified in further studies with larger cohorts.

Reproductive toxicity of microplastics in female mice and their offspring from induction of oxidative stress

Microplastics (MPs) are an emerging pollutant that is becoming recognized as an increasingly serious environmental problem. The biological toxicity and resulting health risks of MPs have attracted much attention in the research community. While the effects of MPs on various mammalian organ systems have been described, their interactions with oocytes and the underlying mechanism of their activity within the reproductive system have remained ambiguous. Here, we discovered that oral administration of MPs to mice (40 mg/kg per day for 30 days) significantly reduced the oocyte maturation and fertilization rate, embryo development, and fertility. Ingestion of MPs significantly increased the ROS level in oocytes and embryos, leading to oxidative stress, mitochondrial dysfunction, and apoptosis. Moreover, mouse exposure to MPs caused DNA damage in oocytes, including spindle/chromosome morphology defects, and downregulation of actin and Juno expression in mouse oocytes. In addition, mice were also exposed to MPs (40 mg/kg per day) during gestation and lactation to determine trans-generational reproductive toxicity. The results showed that maternal exposure to MPs during pregnancy resulted in a decline in birth and postnatal body weight in offspring mice. Furthermore, MPs exposure of mothers markedly reduced oocyte maturation, fertilization rate, and embryonic development in their female offspring. This investigation provides new insights on the mechanism of MPs' reproductive toxicity and raises concerns for potential risks of MP pollution on the reproductive health of humans and animals.

Letrozole cotreatment improves the follicular output rate in high-body-mass-index women with polycystic ovary syndrome undergoing IVF treatment.

Women who have polycystic ovary syndrome (PCOS) with high body mass index (BMI) typically have an attenuated ovarian response and decreased follicular size, which are linked to unfavourable clinical outcomes following in vitro fertilization (IVF) therapy. The follicular output rate (FORT), a qualitative indicator of follicular response, seems to be positively linked to the clinical outcomes of IVF. Progestin-primed ovarian stimulation (PPOS) has become an alternative to gonadotropin-releasing hormone (GnRH) analogues to inhibit the premature luteinizing hormone (LH) surge. As letrozole (LE) shows promise in enhancing ovarian response, we compared PPOS with and without LE for PCOS in high BMI women with a focus on the FORT and associated clinical and pregnancy outcomes. For the recruited 1508 women, ten variables including AFC; age; basal sex hormone level; BMI; infertility type; period of infertility and number of previous IVF attempts were chosen in the propensity score matching (PSM) model to match 1374 women who taken the MPA+ hMG protocol with 134 women who received the MPA+ hMG+ LE treatment at a 1:1 ratio. FORT was selected as the primary outcome measure. The number of oocytes retrieved, viable embryos, hMG dosage, duration, oocyte maturity rate, fertilization rate, and implantation rate were established as secondary outcomes. FORT was substantially elevated in the MPA+hMG+LE group compared with the MPA+hMG group (61% [35%, 86%] vs. 40% [25%, 60%], P <.001). Interestingly, the LE cotreatment group had a considerably lower mature oocyte rate despite having a similar number of mature oocytes and embryos recovered. The average hMG dosages and durations in the study group were similar to those in the control group. The implantation rate in the study group was numerically higher but without statistic significant than that in the control groups (43.15% (107/248) vs. 38.59% (115/298), OR 1.008, 95% CI 0.901-1.127; P >.05). The effect of LE combined with PPOS on FORT is better than the effect of the standard PPOS treatment in women with PCOS and a high BMI, but there is no substantially beneficial impact on pregnancy outcomes or the cycle features of COS, including consumption of hMG.

Astaxanthin treatment ameliorates ER stress in polycystic ovary syndrome patients: a randomized clinical trial

Astaxanthin (ASX), as a natural carotenoid compound, exists in various types of seafood and microorganisms. It has several possible beneficial therapeutic effects for patients with polycystic ovary syndrome (PCOS). Patients with PCOS also suffer from endoplasmic reticulum (ER) stress. In the present work, it was hypothesized that ER stress could be improved by ASX in PCOS patients. Granulosa cells (GCs) were obtained from 58 PCOS patients. The patients were classified into ASX treatment (receiving 12 mg/day for 60 days) and placebo groups. The expression levels of ER stress pathway genes and proteins were explored using Western blotting and quantitative polymerase chain reaction. To assess oxidative stress markers, follicular fluid (FF) was gained from all patients. The Student’s t test was used to perform statistical analysis. After the intervention, ASX led to a considerable reduction in the expression levels of 78-kDa glucose-regulated protein (GRP78), CCAAT/enhancer-binding protein homologous protein (CHOP), and X-box-binding protein 1 compared to the placebo group, though the reduction in the messenger RNA (mRNA) expression level of activating transcription factor 6 was not statistically significant. However, ASX significantly increased the ATF4 expression level. GRP78 and CHOP protein levels represented a considerable decrease in the treatment group after the intervention. In addition, a statistically significant increase was found in the FF level of total antioxidant capacity in the treatment group. Based on clinical outcomes, no significant differences were found between the groups in terms of the oocyte number, fertilization rate, and fertility rate, but the ASX group had higher rates of high-quality oocytes, high-quality embryo, and oocyte maturity compared to the placebo group. Our findings demonstrated that ER stress in the GCs of PCOS patients could be modulated by ASX by changing the expression of genes and proteins included in the unfolding protein response.Trial registration This study was retrospectively registered on the Iranian Registry of Clinical Trials website (www.irct.ir; IRCT-ID: IRCT20201029049183N, 2020-11-27).

Nuclear Maturation Competence and Pronuclear Formation in Ovine Oocytes Supplemented with L-ergothioneine in Maturation Medium

This study aimed to determine the efficacy of supplementing maturation medium with L-ergothioneine on nuclear maturation competence and pronuclei formation of ovine oocytes in vitro.  In the first study, oocytes were divided into four groups and matured for 24 hours in a maturation medium supplemented with 0 mM (control), 10 mM, 15 mM, and 20 mM LE.  In the second study, oocytes were divided into two groups and matured for 24 hours in a maturation medium supplemented with 0 mM LE (control) and 10 mM LE (optimal dose from the first study), then co-incubated with sperm in a fertilization medium for 12 hours. The results of the first study showed that the maturation rate of oocytes enriched with LE 10 mM (88.74±1.79%) and 15 mM (87.52±2.30%) in maturation medium was significantly different (P0.05) compared to the control (78.55±1.78%) and LE 20 mM (77.37±1.21%). Furthermore, the results of the second study revealed that the fertilization rate of oocytes with 10 mM LE (75.55±3.47%) supplementation in maturation medium had a significantly higher percentage of two pronuclei formation than control (63.78±3.49%). In conclusion, supplementing 10 mM LE in a maturation medium improved oocyte nuclear maturation competence and fertilization rates.

Urinary concentration of selected nonpersistent endocrine disrupting chemicals—reproductive outcomes among women from a fertility clinic

Parabens and benzophenones are compounds widely used in cosmetics and personal care products. Although human exposure is widespread there is a limited number of epidemiological studies assessing the relationship between exposure to these chemicals and female reproductive health. The aim of the study is to explore the relationship between paraben and benzophenone concentrations and reproductive outcomes among women attending a fertility center. This prospective cohort included 450 women undergoing in vitro treatment (IVF) at fertility clinic in Poland. The validated gas chromatography ion-tap mass spectrometry to assess concentrations of parabens in urine (methyl (MP), ethyl (EP), propyl (PP), butyl paraben (BP)) and benzophenone-3 (BP-3) was used. To explore the relationship between concentrations of examined chemicals and reproductive outcomes (methaphase II (MII) oocyte yield, total oocyte yield, implantation rate, fertilization rate, clinical pregnancy, live births), multivariable generalized linear mixed model was used for the analysis. Increased exposure to butyl paraben was associated with a significant decrease in MII oocyte count (p = 0.007) when exposure to BP was treated as the continuous variable. Additionally, the exposure to BP in the highest quartile of exposure also decreases MII oocyte count (p = 0.02) compared to the lowest quartile. Urinary concentrations of BP were not related to total oocyte count, fertilization and implantation rate, clinical pregnancy, and live birth when the exposure variable was continuous variable or in the quartiles of exposure. Exposure to MP, EP, PP, the sum of examined parabens, and benzophenone-3 were not related to any of the examined reproductive outcomes. Exposure to butyl paraben was associated with a decrease in MII oocyte count among women attending fertility clinic rinsing concerns that exposure may have a potential adverse impact on embryological outcomes. The results emphasize the importance to reduce chemicals in the environment in order to minimize exposure. As this is the first study showing such an association, further research is needed to confirm these novel results in other populations.Graphical

Thyroid Autoimmunity Is Not Associated with Embryo Quality or Pregnancy Outcomes in Euthyroid Women Undergoing Assisted Reproductive Technology in China.

Background: Studies have shown that thyroid autoimmunity (TAI) is associated with increased risks of adverse pregnancy outcomes. The aim of this study was to investigate the associations between TAI and embryo quality in euthyroid women undergoing in vitro fertilization or intracytoplasmic sperm injection (IVF/ICSI). Methods: This retrospective cohort study included euthyroid infertile women with and without TAI (defined as a serum thyroperoxidase concentration ≥34 IU/mL or a thyroglobulin concentration ≥115.0 IU/mL) who underwent their first complete IVF/ICSI treatment cycles at a tertiary referral center between April 2016 and February 2022. Embryo quality measurements and clinical outcomes were compared between women with (TAI positive) and without TAI (TAI negative). The high-quality cleavage embryo rate and cumulative live birth rate (cLBR) were the primary outcomes. Results: A total of 499 TAI-positive and 2945 TAI-negative women were included in this study, and their mean (standard deviation) ages were 31.6 (4.5) and 30.9 (4.4) years, respectively (p = 0.001). The overall analysis showed no significant differences between TAI-negative and TAI-positive women in the high-quality cleavage embryo rate (n/N: 11,139/22,553 vs. 1971/3820; adjusted rate: 52.8% vs. 53.4%, p = 0.66) and cLBR (1917/2945 vs. 327/499; 53.4% vs. 56.2%, p = 0.31). Moreover, no significant differences were observed between TAI-negative and TAI-positive women in the rates of oocyte retrieval (35,078/51,978 vs. 5853/8628; 69.1% vs. 69.4%; p = 0.65), fertilization (23,067/34,197 vs. 3902/5728; 61.1% vs. 62.2%, p = 0.34), embryo utilization (18,233/22,553 vs. 3156/3820; 80.2% vs. 80.8%, p = 0.61), blastocyst formation (7051/13,721 vs. 1192/2330; 48.5% vs. 48.4%, p = 0.97), and high-quality blastocysts (4819/13,721 vs. 799/2330; 29.9% vs. 29.4%, p = 0.73). Furthermore, no significant differences were observed between TAI-negative and TAI-positive women in the clinical pregnancy rate (1524/2808 vs. 248/482; 46.7% vs. 44.6%, p = 0.40), early pregnancy loss rate (156/1524 vs. 23/248; 13.5% vs. 11.5%, p = 0.44), and LBR (1338/2808 vs. 218/482; 37.4% vs. 36.0%, p = 0.55) of the first transfer cycle. Conclusions: This study demonstrated that TAI in women was not associated with embryo quality or the cLBR following IVF/ICSI. Future large studies are warranted to confirm these findings.

Influence of hypoxanthine concentrations on capacitation, acrosome reaction and In vitro fertilization in Ram Spermatozoa

The aim of the study was to evaluate the effect of hypoxanthine concentrations and incubation time on motility, hyperactivity (HA) and acrosome reaction (AR) in ram sperm in vitro. Semen samples (n= 40) were collected using artificial vagina from 3 mature Barki rams. Freshly ejaculated spermatozoa were pooled and subjected to swim up technique in modified sperm Tyrode's albumin lactate pyruvate (S-TALP) medium supplemented with different concentrations of hypoxanthine (5, 10, 15, 20 mg/ml). Motility, hyperactivity percentage and acrosomal status were recorded at 0, 1, 2, 3 and 4 h post-incubation. They were examined for ability to fertilize oocyte in vitro. The results showed that hypoxanthine concentrations and incubation time for 2 h did not significantly affect the motility. However, increase in incubation time for >2 h significantly (P<0.05) decreased the motility of spermatozoa as compared to control. Treatment of spermatozoa with 20 mg/ml hypoxanthine significant (P<0.05) increased in HA after 2 h of incubation compared to control (5.2±0.3 vs. 1.0± 0.1%). Significant (P<0.05) increase in total AR was associated to the increase in concentration of hypoxanthine with maximum at 20 mg/ml at 3h of incubation (82.0±2.2%). The fertilization rate of oocytes treated with 20 mg/ml hypoxanthine was higher as compared with control (42.8 vs. 26.0%). In conclusion, treatment of ram spermatozoa with 20 mg/ml hypoxanthine for 2 to 3 h was considered the best concentration for in vitro fertilization in sheep.

Impact of the number of retrieved oocytes on IVF outcomes: oocyte maturation, fertilization, embryo quality and implantation rate

The process of oocyte retrieval represents a key phase during the cycles of in vitro fertilization (IVF). It involves controlled ovarian stimulation to retrieve the highest number of oocytes possible. According to many previous studies, the higher the number of oocytes the higher the chances of obtaining embryos for multiple transfers. In this study, in total, 1987 patients were retrospectively reviewed to investigate the correlations between the number of retrieved oocytes and the subsequent IVF outcomes. Patients were divided into three groups according to the number of retrieved oocytes (Group 1: ≤5 oocytes; Group 2: 6-15 oocytes; Group 3: ≥15 oocytes). The results showed a significant negative correlation between oocyte number and maturation rate as well as fertilization rate. However, a significant positive correlation was found between oocyte number and the blastulation rate. The implantation rate after fresh embryo transfers was higher in group 2 (6-15 oocytes) compared with group 1 (≤5 oocytes). According to our findings, we conclude that oocyte numbers between 6 and 15 oocytes can result in the highest chances of positive IVF outcomes in terms of embryo quality and fresh embryo transfers with lower risks of ovarian hyperstimulation.

Progestin-primed ovarian stimulation protocol with or without clomiphene citrate for poor ovarian responders: a retrospective cohort study

ObjectiveTo explore the efficacy of progestin-primed ovarian stimulation (PPOS) combined with clomiphene citrate (CC) versus PPOS protocol used alone on cycle characteristics and pregnancy outcomes for women with the poor ovarian response (POR).MethodsWe performed a retrospective cohort study and a total of 578 POR patients who underwent IVF/ICSI cycles were collected and divided into Group A (HMG 300 IU/d + MPA 10 mg/d) and Group B (HMG 300 IU/d + MPA 10 mg/d + CC 50 mg/d). The primary outcome measure was the number of oocytes retrieved, other outcome measures were cycle characteristics and clinical pregnancy rate.ResultsThe baseline information between the two groups were not statistically significant (P > 0.05). Compared with Group A, Group B had a lower total dose of human menopausal gonadotrophin (HMG) (2998.63 ± 1051.09 vs. 3399.18 ± 820.75, P < 0.001) and the duration of stimulation (10.21 ± 3.56 vs. 11.27 ± 2.56, P < 0.001). Serum luteinizing hormone level was higher in Group B on human chorionic gonadotrophin injection day (P < 0.001). The number of oocyte for retrieval, maturation, and fertilization were significantly lower in Group B than that in Group A (P < 0.001). However, the oocyte retrieval rate, maturation rate, fertilization rate, and viable embryo rate showed no statistical difference in the two groups (P > 0.05). After adjusting for confounders, the clinical pregnancy rate (OR 1.286; 95% CI 0.671–2.470) and live birth rate (OR 1.390; 95% CI 0.478–3.990) were comparable between the two groups.ConclusionsPPOS protocol combined with CC reduces the total dose of HMG and the duration of stimulation, and can also achieve similar oocyte yields and clinical pregnancy rate compared with the PPOS protocol used alone in poor ovarian responders.

EVALUATION OF OOCYTE FERTILIZATION AND IN VITRO EMBRYO DEVELOPMENT AFTER SPERM PREPARATION USING ZYMŌT® PRIOR TO INTRACYTOPLASMIC SPERM INJECTION (ICSI) AND IN VITRO FERTILIZATION (IVF)

To compare ICSI/IVF oocyte fertilization rates and in vitro embryo development associated with the use of ZyMōt®, versus conventional semen preparation methods, prior to selecting sperm for injection.

Follicular flushing increases the number of oocytes retrieved in women with diminished ovarian reserve and serum progesterone levels of ≤ 5.0 ng/ml post surge

ObjectiveTo evaluate the utility of follicular flushing increase the number of oocytes retrieved in women with diminished ovarian reserve (DOR) and serum progesterone of ≤ 5.0 ng/ml the day after final oocyte maturation induction.DesignRetrospective study.SubjectsWomen &lt;4O years, with DOR, underwent controlled ovarian stimulation for an invitro fertilization cycle (IVF) were separated into two groups: Group 1: follicular flushing (n=35) Group 2: direct aspiration (n=33).Main outcomes measuredOur primary outcome was the number of retrieved oocytes. Secondary outcome measures included the total procedure time, mean oocyte/follicle retrieval rate, mean number of metaphase II (MII) oocytes, MII rate, mean number of fertilized oocytes, and blastulation rate.ResultsA total of 288 oocytes were retrieved: 59.0% from the flushing group and 41.0% from the direct aspiration group. The mean number of oocytes retrieved by aspiration and subsequent flushes was significantly higher than the number retrieved from direct aspiration (4.4 ± 0.96 versus 2.64 ± 1.00, P = 0.003). The oocyte/follicle rate was also significantly higher in women in Group 1 vs Group 2 (68.3% vs 48.7%, p=0.001). No significant differences were observed in the total number of MII, oocyte maturation rate, fertilization rate nor blastulation rate. The retrieval procedure time was higher among those who underwent follicular flushing, with an estimated increase of 6.5 minutes (14.0 ± 2.5 versus 7.5 ±1.7, P = 0.003).&#x0D; ConclusionsThe results of this analysis in the poor responders found that women with DOR and post surge P4 levels of ≤ 5.0 ng/ml may benefit from follicular flushing on the number of oocytes retrieved.

Study on the expression patterns and function of JUNO and CD9 in bovine oocytes during in vitro maturation.

Fertilization proteins JUNO and CD9 play vital roles in sperm-egg fusion, but little is known about their expression patterns during in vitro maturation (IVM) and their function during in vitro fertilization (IVF) of bovine oocytes. In this study, qRT-PCR and immunofluorescence staining were used to detect the mRNA and protein expression levels of JUNO and CD9 genes in bovine oocytes and cumulus cells. Then, fertilization rate of MII oocytes treated with (i) JUNO antibody (1, 5 and 25 μg/ml) or (ii) CD9 antibody (1, 5 and 25 μg/ml) or (iii) CD9 antibody (5μg/ml) + JUNO antibody (5μg/ml) were recorded. Our results showed that the mRNA and protein expression levels of JUNO and CD9 genes significantly increased from bovine GV oocytes to MII oocytes, and similar mRNA expression patterns of JUNO and CD9 were also detected in cumulus cells. All groups of oocytes treated with CD9 antibody or JUNO antibody showed significantly decreased fertilization rates (p < .05). Particularly, the fertilization ability of oocytes treated with CD9 antibody (5μg/ml) + JUNO antibody (5μg/ml) sharply decreased to 3.48 ± 0.11%. In conclusion, our study revealed the expression levels of JUNO and CD9 genes in oocytes and cumulus cells increased during IVM of bovine oocytes, with JUNO protein playing a major role in the fertilization of bovine oocytes.

Comparison of oocyte count, fertilization, and pregnancy rates in adenomyosis patients undergoing In Vitro Fertilization with short and long protocol controlled ovarian stimulation - Restospective study.

IntroductionAdenomyosis (benign gynecological disease) is an endometrial stromal tissue condition that invades the myometrium of the uterus. The administration of a gonadotropin-releasing hormone (GnRH) analog before the IVF (In Vitro Fertilization) cycle significantly increases the chances of getting pregnant in adenomyosis patient with infertile condition. There is no best protocol consensus for adenomyosis for now. This study plans to compare the outcomes of long-protocol and short-protocol of IVF in adenomyosis patients who have undergone surgery and treatment with GnRH analogs.MethodThis study is a retrospective study with a comparative method. The study was conducted at the IVF Aster Clinic and the IVF Bandung Fertility Center by reviewing retrospective data from 2015 to 2021. Patients who have been diagnosed with adenomyosis will undergo IVF procedure with a long-protocol and short-protocol pretreatment. Parameters observed were oocyte count, fertilization, and pregnancy rate.ResultsFifty-eight patients were included. There was a significant difference in the oocytes count and the pregnancy rate in short and long groups with p value of less than 0.05, while there was no significant difference in fertilization rate with p value of greater than 0.05.ConclusionThere were differences in the oocytes count and pregnancy rates in the short and long protocol groups in adenomyosis patients. There was no difference in fertilization rate in the short and long protocol groups in adenomyosis patients.

Anti–Müllerian hormone and antral follicle count predict ovarian response in women less than 45 years following GnRH antagonist multiple–dose protocol

Objective: To speculate which of the following parameters: antral follicle count (AFC), anti-Müllerian hormone (AMH), follicle- stimulating hormone (FSH) and age can be used as a predictor of ovarian response to gonadotropin-releasing hormone (GnRH) antagonist stimulation multiple-dose protocol in women under 45 years, and to determine the cutoff value of these parameters and their correlations for predicting low and high ovarian response. Methods: This prospective study included 462 women with the mean age of (29.3±6.5) years. All women were subjected to the GnRH antagonist stimulation multiple-dose protocol. On the second day of the menstrual cycle, ultrasonography was conducted to determine AFC in both ovaries. Peripheral blood samples were collected to evaluate the level of estradiol, FSH, luteinizing hormone, prolactin, thyroid-stimulating hormone, and AMH. The women were divided into three groups: low response (AHH&lt;1 ng/mL, n=173), normal response (AMH=1.0-3.5 ng/mL, n=175), and high response (AMH &gt;3.5 ng/mL, n=114). Results: A significant decrease was found in the age and FSH level in the high response group compared to other groups (P&lt;0.001). Conversely, a significant increase was shown in AMH, estradiol on human chorionic gonadotropin (hCG) day, AFC, mature oocytes, fertilized oocytes, and embryos transferred in the high response group compared to the other two groups (P&lt;0.001). The receiver operating characteristic (ROC) curves demonstrated that AFC and AMH had the highest accuracy, followed by basal FSH level and age in the prediction of low ovarian reserves (P&lt;0.001) with cutoff values of ≤4.50 and ≤0.95 for AFC and AMH, respectively. Moreover, the ROC analysis showed that AFC had the highest accuracy, followed by AMH level and age in the prediction of high ovarian reserves with a cutoff value of ≥14.50, ≥3.63, and ≤27.50 years, respectively (P&lt;0.01). A significant decrease was observed in women's age, estradiol level, and oocyte fertilization rate in pregnant women compared to non-pregnant women (P&lt;0.001). Additionally, significant negative correlations were found between the AFC, the number of mature oocytes, fertilized oocytes, embryos transferred, and the age of pregnant women (P&lt;0.001). Conclusions: AFC and AMH predict low and high ovarian response to GnRH antagonist stimulation multiple-dose protocol in women under 45 years.

Implementation and Evaluation of Preimplantation Genetic Testing at Vilnius University Hospital Santaros Klinikos.

The most effective treatment of infertility is in vitro fertilization (IVF). IVF with Preimplantation Genetic Testing (PGT) allows to identify embryos with a genetic abnormality associated with a specific medical disorder and to select the most optimal embryos for the transfer. PGT is divided into structural rearrangement testing (PGT-SR), monogenetic disorder testing (PGT-M), and aneuploidy testing (PGT-A). This study mostly analyzes PGT-SR, also describes a few cases of PGT-M. The aim of this study was to implement PGT procedure at Vilnius University Hospital Santaros Klinikos (VUHSK) Santaros Fertility Centre (SFC) and to perform retrospective analysis of PGT procedures after the implementation. A single-center retrospective analysis was carried out. The study population included infertile couples who underwent PGT at SFC, VUHSK from January 01st, 2017 to December 31st, 2020. Ion PGM platform (Life Technologies, USA) and Ion ReproSeq PGS View Kit (Life Technologies, USA) were used for the whole genome amplification. Results were assessed using descriptive statistics. PGT was successfully implemented in VUHSK in 2017. During the analyzed time period, thirty-four PGT procedures were performed for 26 couples. Two procedures were performed in 2017, 7 procedures - in 2018, 13 - in 2019, and 12 - in 2020. In comparison with all IVF procedures, 2.5% procedures were IVF with PGT, a highest percentage was in 2020 (3.8% of all procedures). The main indication for PGT was balanced chromosomal rearrangements (in 85.3% cases). In all 34 cases 515 oocytes were aspirated in total, 309 oocytes were fertilized, oocytes fertilization rate exceeded 60%. A normal diploid karyotype was found in 46 (16.8%) biopsied embryos. Out of all PGT procedures, 9 (26.5%) resulted in a clinical pregnancy. Six (66.7%) pregnancies were confirmed in 2019, and 3 (33.3%) - in 2020. Three (33.3%) pregnancies resulted in spontaneous abortion, 6 (66.7%) - in delivery. The implementation of PGT in VUHSK was successful. The most common indication for PGT was a reciprocal translocation. Oocytes fertilization rate exceeded 60%, a normal karyotype was found less than in one-fifth of biopsied embryos. A highest clinical pregnancy rate was achieved in 2019 when almost half of women conceived, which is probably related to the experience gained by the multidisciplinary team. This is the first study analyzing IVF with PGT in Lithuania, however, the results should be interpreted with caution due to a low number of total procedures performed.

Letrozole cotreatment with progestin-primed ovarian stimulation in women with polycystic ovary syndrome undergoing IVF treatment.

Background: Progestin is an alternative to gonadotropin-releasing hormone (GnRH) analogues in the follicular phase to suppress the premature luteinizing hormone (LH) surge in women with polycystic ovary syndrome (PCOS). However, progestin-primed ovarian stimulation (PPOS) is always accompanied by increased pituitary suppression and gonadotropin consumption. Previous studies suggested that letrozole appeared to have the potential to reduce the total gonadotropin dose required for ovarian stimulation. A retrospective cohort study was performed to evaluate the efficacy of PPOS with or without letrozole in infertile women with PCOS. Methods: This retrospective cohort study included 448 women with PCOS who underwent controlled ovarian stimulation (COS) with human menopausal gonadotropin (hMG) and medroxyprogesterone acetate (MPA) (n = 224) or hMG and MPA cotreatment with LE (n = 224) from January 2018 to March 2021 after propensity-score matching. The primary outcome measure was the hMG dose. The secondary outcomes were the durations of ovarian stimulation, the implantation rate, the number of oocytes retrieved and viable embryos, oocyte maturity and fertilization rates, the percentage of women with profound pituitary suppression (luteinizing hormone [LH] <1.0 IU/L on the trigger day). Results: The hMG doses (1949.89 ± 725.03 IU vs 2017.41 ± 653.32 IU, p > 0.05) and durations of ovarian stimulation (9.03 ± 1.79 days vs 9.21 ± 2.18 days, p > 0.05) were similar between the two groups. The implantation rate was significantly higher in the study group (MPA + hMG + LE) than in the control group (MPA + hMG) (42.22 vs 34.69%, p < 0.05). The numbers of oocytes and embryos retrieved were similar between the two groups. Interestingly, letrozole cotreatment was associated with decreased oocyte maturity and fertilization rates in comparison with standard PPOS protocols even though mature and fertilized oocyte yields were comparable. Compared with those in the control group, the LH values on the trigger day were significantly higher in the study group, together with significantly reduced pituitary suppression. Conclusion: Letrozole combined with PPOS cannot reduce hMG consumption in PCOS patients undergoing IVF treatment and shows no beneficial effect on cycle characteristics of COS. However, letrozole supplementation manifests as a superior implantation rate to that of the standard PPOS protocol in women with PCOS.

P-148 Denudation employing human recombinant hyaluronidase enzyme does not affect fertilization and blastocyst formation rates in sibling oocytes from patients undergoing ART: A prospective randomized study

Abstract Study question Does human recombinant hyaluronidase (Cumulase®) improve fertilization and blastocyst formation rates in sibling human oocytes compared to bovine-derived hyaluronidase? Summary answer The use of Cumulase® for oocyte denudation does not appear to affect fertilization or usable blastocyst formation rate in patients undergoing intracytoplasmic sperm injection (ICSI). What is known already Bovine-derived hyaluronidase has been widely employed to denude the cumulus-oocyte complexes (COCs) prior to ICSI or oocyte vitrification. Nonetheless, the recent concern for its safety, due to its animal origin, has led to the development of alternatives ascertaining the enzyme’s purity. It has been reported that human recombinant hyaluronidase is not equally effective as its bovine-derived analogue, leading to a longer time required for complete denudation coupled by the subsequent toxicity risk for oocytes. This may result to lower fertilization and blastocyst formation rates. Although there are some studies on IVF outcomes with recombinant hyaluronidase, its use remains rather limited. Study design, size, duration This randomized double-blinded clinical trial was conducted at Citmer Reproductive Medicine, in Puebla and Monterrey, Mexico from February to December 2021. The impact of denudation on sibling oocytes from 103 patients employing bovine-derived hyaluronidase (InVitroCare®) or human recombinant hyaluronidase (ICSI Cumulase® (rHuPH20), CooperSurgical®) was evaluated. A total of 1237 oocytes were assigned as follows: The Hyaluronidase Group included 626 oocytes, while the Cumulase® Group included 611 oocytes. Preimplantation Genetic Testing was performed in 18 cycles. Participants/materials, setting, methods After retrieval, COCs were incubated for 2-4 h and placed in microdropets with 10 IU/mL bovine-derived hyaluronidase or Cumulase® in culture media GlobalTotal® (LifeGlobal®) for granulosa cells removal. COCs were gently pipetted up and down employing 20 μL and 150 μm tips. Metaphase II oocytes were injected and incubated for 17-20 hours in 8% CO2, 20% O2, 37°C. Normally fertilized zygotes were cultured to blastocyst stage and embryo development was evaluated. Seventy-nine blastocysts were biopsied. Main results and the role of chance The mean age of the patients was 32.12±1.11 years old. A total of 506 and 493 metaphase II oocytes from Groups 1 and 2 respectively were subjected to ICSI. Normal fertilization rates for oocytes treated with Hyaluronidase and Cumulase® were 71.34% (n = 361) and 72.41% (n = 357) respectively, with no statistically significant differences between the two groups (p = 0.35). Usable blastocysts rate (selected for embryo transfer or blastocyst cryopreservation) did not differ with any statistical difference between the bovine-derived hyaluronidase (n = 170, 47.09%) and the Cumulase® treated oocytes (n = 158, 44.25%), (p = 0.44). Preimplantation genetic testing for aneuploidies (PGT-A) was performed in 18 cycles. Seventy-nine blastocysts were biopsied and tested. Fourty-seven embryos from the bovine-derived hyaluronidase group were biopsied, presenting with a euploidy rate of 51.06% (n = 24), compared to 32 embryos from the recombinant hyaluronidase group with a euploidy rate of 50% (n = 16). Results indicated no statistically significant difference (p = 0.92). These data support that human recombinant hyaluronidase presents with similar efficacy to bovine-derived hyaluronidase. in terms of normal fertilization, embryo development as well as euploidy rates. Limitations, reasons for caution The small sample size regarding the euploidy outcome stands as a limitation of the study. Further studies should focus on the effect of recombinant hyaluronidase on genetic anomalies on blastocysts. Wider implications of the findings Fertilization, usable blastocyst and euploidy rates did not differ when employing bovine-derived or human recombinant hyaluronidase, rendering the latter a safe and efficient alternative. However, it is important to assess long-term effects, as only a limited number of studies report on perinatal and neonatal outcomes Trial registration number 25-12-21MTYPUECIT