Normal Fertilization Rate Research Articles

Intracytoplasmic sperm injection compared with in vitro fertilisation in patients with non-male factor infertility with low oocyte retrieval: a single-centre, retrospective cohort study

ObjectiveTo investigate the effects of in vitro fertilisation (IVF) and intracytoplasmic sperm injection (ICSI) on the clinical outcomes of non-male factor patients aged >35 with three or fewer oocytes retrieved.DesignRetrospective...

Effects of ovarian stimulation on embryo euploidy: an analysis of 12874 oocytes and 3106 blastocysts in cycles with preimplantation genetic testing for monogenic disorders.

Does ovarian stimulation and the ovarian response affect embryo euploidy? Ovarian stimulation and the ovarian response in women undergoing preimplantation genetic testing for monogenic disorders (PGT-M) cycles did not affect the rates of blastocyst euploidy. Whether or not ovarian stimulation in IVF-embryo transfer has potential effects on embryo euploidy is controversial among studies for several reasons: (i) heterogeneity of the study populations, (ii) biopsies being performed at different stages of embryo development and (iii) evolution of the platforms utilized for ploidy assessment. Patients who undergo PGT-M cycles typically have no additional risks of aneuploidy, providing an ideal study population for exploring this issue. A retrospective cohort study including embryos undergoing PGT-M was conducted at a single academically affiliated fertility clinic between June 2014 and July 2021. A total of 617 women with 867 PGT-M cycles involving 12874 retrieved oocytes and 3106 trophectoderm biopsies of blastocysts were included. The primary outcome of the study was median euploidy rate, which was calculated by dividing the number of euploid blastocysts by the total number of biopsied blastocysts for each cycle. Secondary outcomes included the median normal fertilization rate (two-pronuclear (2PN) embryos/metaphase II oocytes) and median blastulation rate (blastocyst numbers/2PN embryos). Comparable euploidy rates and fertilization rates were observed across all age groups, regardless of variations in ovarian stimulation protocols, gonadotropin dosages (both the starting and total dosages), stimulation durations, the inclusion of human menopausal gonadotrophin supplementation, or the number of oocytes retrieved (all P > 0.05). Blastulation rates declined with increasing starting doses of gonadotropins in women aged 31-34 years old (P = 0.005) but increased with increasing gonadotrophin starting doses in women aged 35-37 years old (P = 0.017). In women aged 31-34, 35-37, and 38-40 years old, blastulation rates were significantly reduced with increases in the number of oocytes retrieved (P = 0.001, <0.001, and 0.012, respectively). Limitations include the study's retrospective nature and the relatively small number of patients of advanced age, especially patients older than 40 years old, leading to quite low statistical power. Second, as we considered euploidy rates as outcome measures, we did not analyze the effects of ovarian stimulation on uniform aneuploidy and mosaicism, respectively. Finally, we did not consider the effects of paternal characteristics on embryo euploidy status due to the fact that blastocyst aneuploidy primarily originates from maternal meiosis. However, sperm factors might have an effect on embryo development and the blastulation rate, and therefore also the number of blastocysts analyzed. The exclusion of patients with severe teratozoospermia and the fact that only ICSI was used as the insemination technique for women undergoing PGT-M contributed to minimize the effect of paternal factors. Ovarian stimulation and response to stimulation did not affect blastocyst euploidy rates in women undergoing PGT-M cycles. However, in women aged 31-40 years old, there was a significant decline in blastulation rates as the number of retrieved oocytes increased. This study was supported by the National Natural Science Foundation of China (Grant No. 81701407, 82301826); the National Key Research and Development Program of China (2022YFC2702901, 2022YFC2703004); China Postdoctoral Science Foundation (2022M710261), and China Postdoctoral Innovation Talent Support Program (BX20220020). There is no conflict of interest. N/A.

Comparative analysis of short versus long co-incubation of gametes on post-insemination outcomes and embryo morphokinetics: a sibling oocyte randomized study

Research questionDoes a short co-incubation of gametes in conventional IVF impact post-insemination outcomes and embryo morphokinetics? DesignSibling oocyte randomized pilot study performed between December 2020 and March 2023. Eligible couples (n=55) were female aged 18-43 years with body mass index (BMI) ≤35 and male normal semen parameters. Cumulus oocyte complexes (COCs) (min 6 - max 12) were randomized in a 1:1:2 proportion in long (16-18 hrs), or short (2 hrs) co-incubation IVF exposure, and ICSI, respectively. All oocytes inseminated via IVF were monitored by time-lapse (TL). Blastocysts of ≥BL3CC were subjected to trophectoderm (TE) biopsy on day (D)5, D6 or D7 by next generation sequencing (NGS). ResultsA total of 954 COCs were distributed to long co-incubation (n=235, 24.6%), short co-incubation (n=235, 24.6%), and ICSI (n=484, 50.3%). In total, 202 were MII after long co-incubation, 209 after short co-incubation and 394 at ICSI. Regression analyses showed both short co-incubation groups (OR: 0.72, 95% CI: 0.46–1.11, P=0.139) and ICSI (OR: 1.43, 05% CI: 0.95–2.15, P=0.090) did not significantly impact normal fertilization rates (2PN) when compared to long co-incubation. Usable blastocysts per MII, per 2PN, and euploid blastocysts per MII, per 2PN and per tested blastocysts were similar among the groups. The rate of embryo arrest (HR: HR: 0.79, 95% CI: 0.57-1.11, P=0.183) and timings to reach morphokinetic milestones among embryos reaching blastulation were similar between IVF groups (P>0.05, for all). ConclusionA short 2-hour co-incubation of gametes in IVF yields comparable fertilization, blastocyst, and euploidy rates without adverse embryo morphokinetic events, compared to standard overnight co-incubation.

Blastocysts originated from oocytes with smooth endoplasmic reticulum aggregates have a reduced euploidy rate: a retrospective cohort study.

Does the presence of smooth endoplasmic reticulum aggregates (SERa) in oocytes adversely impact the euploidy rate of subsequent blastocysts? We performed a retrospective cohort study with 671 young patients (< 38 years) undergoing their first preimplantation genetic testing for aneuploidy (PGT-A) between January 2019 and October 2022 at a reproductive medical center of university affiliated teaching hospitals in China. Cycles were categorized as either SERa(+) cycles (containing at least one SERa(+) oocyte) or SERa(-) cycles (all oocytes without SERa). In SERa(+) cycles, oocytes were further subdivided into the SERa(+) oocyte group and the sibling SERa(-) oocyte group, comprising oocytes with normal morphology. No significant differences were observed in the normal fertilization rate (72.9% vs. 75.4% vs. 72.6%, P=0.343), and cleavage rate (96.8% vs. 97.1% vs. 96.4%, P=0.839) among the SERa(-) cycle group, the SERa(-) oocyte group, and the SERa(+) oocyte group. Additionally, there were no statistically significant differences in the rates of good quality embryos (44.7% vs. 48.8% vs. 46.2%, P=0.177) or blastocyst formation (60.1% vs. 60.9% vs. 60.5%, P=0.893) among the groups. However, the euploidy rate of blastocysts derived from SERa(+) oocytes was significantly lower compared to those from SERa(-) oocytes in SERa(+) cycles and normal oocytes in SERa(-) cycles (39.3% vs. 51.2% vs. 54.5%, P=0.005). Despite this, there were no significant differences in pregnancy and neonatal outcomes after euploid embryo transfer among the three groups. Blastocysts derived from SERa(+) oocytes have a lower euploidy rate than those derived from SERa(-) oocytes. Nevertheless, comparable reproductive outcomes were achieved following euploid embryo transfer from both SERa(+) and SERa(-) oocytes.

Rescue ICSI Improved Cumulative Live Birth Rate for Cycles with Second Polar Body Extrusion Rate

ObjectiveTo determine the indications for early rescue intracytoplasmic sperm injection (ICSI) application. DesignA retrospective cohort study SubjectsThe study included 19,808 patients who underwent conventional in vitro fertilization (IVF) or rescue ICSI for their first cycles between February 2017 and December 2021. ExposureRescue ICSI cycles constituted the study group, where oocytes that had not extruded the second polar body 4-6 hours after insemination were rescued by ICSI. The control group consisted of conventional IVF cycles with no interventions to rescue oocytes without the second polar body. Generalized additive models (GAMs) were constructed to describe the relationship between the second polar body extrusion rate and cumulative live birth rate in conventional IVF and rescue ICSI cycles, respectively. The cutoff value of the second polar body extrusion rate guiding rescue ICSI application was determined from the intersection point of GAMs. Maternal age range applicable to rescue ICSI was further analyzed using the same method. Clinical outcomes were compared between conventional IVF and rescue ICSI cycles across different second polar body extrusion rate and maternal age subgroups. Main outcome measuresThe second polar body extrusion rate and maternal age range for rescue ICSI application, normal fertilization rate, and cumulative live birth rate. ResultsGAMs showed that the cutoff value for the second polar body extrusion rate about rescue ICSI application was 50%. When the rate <50%, normal fertilization rate and cumulative live birth rate (63.7% vs. 46.1%, OR: 1.609, 95% CI: 1.276-2.030) were significantly higher in rescue ICSI cycles than conventional IVF cycles. When the rate ≥50%, rescue ICSI cycles had similar normal fertilization rate and cumulative live birth rate compared to conventional IVF cycles. Further analysis on maternal age in cycles with second polar body extrusion rate <50% released that rescue ICSI cycles showed a higher cumulative live birth rate (67.7% vs. 48.3%, OR: 1.732, 95% CI: 1.361-2.202) than conventional IVF cycles for women aged < 38 years. ConclusionIVF cycles with second polar body extrusion rate <50% in women aged < 38 years was applicable to early rescue ICSI.

High risk of low birth weight in couples with advanced paternal age in in vitro fertilization treatment.

With the development of socio-economic conditions and a shift in attitudes towards fertility, there has been a gradual increase in delayed childbearing since the 2000s. Ageplays a significant role in the decline of fertility. However, we know very little about the association of paternal age with reproductive outcomes. To investigate the correlation between advanced paternal age and semen quality, embryo quality, pregnancy, and neonatal outcomes in IVF cycles. In this study, after excluding female partners aged ≥35 years, we analyzed data from 761 infertile couples who underwent in vitro fertilization cycles at the First Affiliated Hospital of USTC between June 2020 and March 2023. Cases were classified into three groups according to the age of the male: <35 years (530 infertile couples), 35 years ≤ paternal age <40 years (125 infertile couples), and ≥40 years (106 infertile couples). Then, we compared the general clinical data arising from in vitro fertilization cycles between the three groups, including semen parameters, embryonic parameters, and pregnancy and neonatal birth outcomes. Data analysis showed that the duration of infertility and the incidence of secondary infertility were significantly higher in paternal age ≥35 years groups than those aged <35 years (all p<0.05). We also observed a significant difference between ≥40 years and <35 years groups in terms of the normal fertilization rate, high-quality embryo rate, clinical pregnancy rate, miscarriage rate, live birth rate, Apgar scores, and the low birth weight neonatal rate (all p<0.05). The group with paternal age ≥40 years showed statistically significant differences in terms of clinical pregnancy rate, miscarriage rate, live birth rate, and low birth weight on multivariable logistic regression (all p<0.05). The results of our study indicate that advanced paternal age (≥40 years) has a significant impact on the embryo quality, pregnancy outcome, and neonatal outcome. Paternal age over 40 years is a risk for in vitro fertilization success rate.

Positive Effect of 25-Hydroxyvitamin D (25(OH)D) Levels in Follicular Fluid on Embryonic Developmental Potential in Diminished Ovarian Reserve (DOR) Patients Undergoing Microstimulation.

This study aimed to explore the effect of 25-hydroxyvitamin D (25(OH)D) levels in follicular fluid (FF) on the embryo outcome with diminished ovarian reserve (DOR) patients undergoing in vitro fertilization (IVF) by microstimulation protocol. A prospective cohort study of 79 patients with DOR who underwent IVF using the microstimulation protocol was conducted. Based on the level of 25(OH)D in follicular fluid (25(OH)D-FF), the patients were divided into a high-value group (25(OH)D-FF>11.1, n = 50) and a low-value group (25(OH)D-FF>11.1, n = 29). Correlation analysis was conducted between the level of 25(OH)D-FF and the rate of high-quality embryos on day 3 (D3). Logistic regression analysis of factors affecting the presence or absence of D3-available embryos in patients with DOR was conducted. The number of oocytes retrieved, mature oocytes (MII), normal fertilization rate, number of available embryos on D3, and high-quality embryo rate were lower in the low-value group than in the high-value group (p = 0.000, p = 0.000, p = 0.009, p = 0.000, p = 0.001). The clinical pregnancy rate of frozen embryo transfer (FET) between the two groups was no statistically significant difference (p > 0.05); correlation analysis between the 25(OH)D-FF level and the rate of high-quality embryos was performed using Spearman's rank-sum test, and there was a positive correlation (R = 0.271, P<0.01). Logistic analysis showed that 25(OH)D-FF level was a protective factor for embryonic outcome (odds ratio (OR) > 1, P<0.01). The 25(OH)D level in FF has a positive effect on embryonic outcomes in DOR patients with IVF using the microstimulation protocol. Vitamin D (VD) supplementation can be used to increase the number of available embryos and improve the quality of embryos for patients with DOR who are undergoing microstimulation of IVF.

A hyaluronic acid-containing reagent compatible with glass-bottom dishes and capable of sustained binding of human spermatozoa.

Commercial products currently available for sperm selection utilizing hyaluronic acid (HA) binding prior to intracytoplasmic sperm injection (ICSI) are widely used but have some disadvantages. To potentially circumvent these limitations, we compared ICSI using a self-made hyaluronic acid (smHA) reagent with ICSI using SpermSlow. The binding of the reagents to spermatozoa on plastic- or glass-bottom dishes was quantitated using spermatozoa that were isolated by density-gradient centrifugation and swim-up procedures (N=10/group). Additionally, we investigated the relationship between the HA reagent used prior to ICSI and clinical outcomes after assisted reproduction with HA-ICSI (N=81). The smHA reagent exhibited extremely stable binding to human spermatozoa. The binding time of spermatozoa was significantly longer in the smHA reagent than in SpermSlow on both plastic and glass dishes (plastic: 60.0±0.0 min vs. 2.7±5.9 min, P<0.001; glass: 60.0±0.0 min vs. 2.5±1.8 min, P<0.001). There were no significant differences in the normal fertilization rate between HA-ICSI with the smHA reagent (128/160, 80.0%) and HA-ICSI with SpermSlow (171/231, 74.0%, P=0.184). The frequency of the blastocyst development from the HA-ICSI-derived zygote was significantly higher with the smHA reagent (74/101, 73.3%) than with SpermSlow (76/131, 58.0%, P=0.019). The rates of biochemical pregnancy, clinical pregnancy, fetal heart movement, live birth, and miscarriage were not significantly different between HA-ICSI with the smHA reagent and HA-ICSI with SpermSlow. The blastulation rate was higher for HA-ICSI with the smHA reagent as compared with SpermSlow. Clinical outcomes, excluding blastulation, after HA-ICSI were the same using smHA reagent and using SpermSlow. Spermatozoa binding to the smHA reagent was not attenuated over a 60-min time course. In conclusion, this reagent may shorten and simplify HA-ICSI procedures because smHA can be used with any dish material, making it easier to observe the spindle or assess intracytoplasmic morphology.

Ambient air pollution decreased normal fertilization rate via the mediation of seminal prosaposin

ObjectiveThis study focuses on the association between seminal concentration of prosaposin and ambient air pollutants and whether the association affects the normal fertilization rate in vitro fertilization (IVF) treatment. MethodsThe cohort of 323 couple participants aged 22–46 was recruited from Jan. 2013 to Jun. 2018. At enrollment, resident address information was obtained and semen parameters of male counterparts were evaluated according to WHO criteria. We used inverse distance weighting interpolation to estimate the levels of ambient pollutants (SO2, O3, CO, NO2, PM2.5, and PM10) in the surrounding area. The exposure of each participant was estimated based on the data gathered from air quality monitoring stations and their home address over various periods (0–9, 10–14, and 0–90 days) before semen sampling. The generalized linear regression model (GLM) and the Bayesian kernel machine regression (BKMR) were used to analyze the associations between pollutants, semen parameters, prosaposin, and normal fertilization. Additionally, the mediating effect of prosaposin and semen parameters on the link between pollutants and normal fertilization was investigated. ResultsGLM and BKMR showed exposure to ambient air pollutants was all associated with the concentration of seminal prosaposin, among them, O3 and CO were also associated with normal fertilization (-0.10, 95 %CI: −0.13, −0.06; −26.43, 95 %CI: −33.79, −19.07). Among the semen parameters, only the concentration of prosaposin and total motile sperm count (TMC) was associated with normal fertilization (0.059, 95 %CI: 0.047, 0.071; 0.016, 95 %CI: 0.012, 0.020). Mediation analysis showed that prosaposin played a stronger mediating role than TMC in the relationship between short-term exposure to O3 and fertilization (66.83 %, P<0.001 versus 3.05 %, P>0.05). ConclusionSeminal plasma prosaposin showed a stronger meditating effect reflect the correlation between ambient air pollutants and normal fertilization rate than conventional semen parameters, which may be used as one of the indicators between pollution and fertilization in IVF.

P-228 Competence of injected MI versus MII sibling oocytes in terms of normal fertilization, blastocyst development and euploidy

Abstract Study question Is the injection of immature MI oocytes as effective as injection of their siblings mature MII oocytes regarding normal fertilization, blastocyst development and euploidy? Summary answer MI oocytes show a lower rate of fertilization, a similar blastulation rate, and a slightly lower rate of euploidy compared to their MII sibling oocytes. What is known already Although several studies have been performed on immature MI oocytes evaluating their developmental competence after intracytoplasmic sperm injection (ICSI), all have been performed on oocytes that extruded the first polar body by the time of injection. ICSI is typically performed on MII oocytes which have completed the process of maturation. Some laboratories also inject MI oocytes, nevertheless, they are generally not used due to the lack of required molecular and cellular changes to support normal development. However, there is limited research on the reproductive potential of oocytes in MI. Consequently, our current practices are founded on assumptions. Study design, size, duration This is a retrospective, observational study of 168 oocytes injected in MI and 839 sibling oocytes injected in MII from 82 ICSI cycles with Preimplantation Genetic Testing for Aneuploidies (PGT-A) performed in a single IVF clinic between April 2018 and December 2023. All PGT-A cycles included in this study contained at least one MI oocyte injected. Participants/materials, setting, methods After oocyte retrieval and denudation, the oocytes were classified as MII, MI or GV. MI oocytes with extruded polar body by the moment of ICSI were reclassified as MII. All MII and MI oocytes were inseminated and the embryos that reached blastocyst stage were biopsied on day 5, 6 or 7. Normal fertilization rate, blastocyst formation rate and euploidy rate were compared between MI-derived embryos and their MII-derived siblings. Main results and the role of chance A total of 1,094 oocytes were retrieved from 83 ICSI cycles with PGT-A, comprising 839 (76.7%) MII, 168 (15.4%) MI y 87 (7.9%) VG. The normal fertilization rate was significantly higher in oocytes injected at MII compared to MI (76.9% vs 53.0%, P &amp;lt; 0.001). The rates of abnormal fertilization in MII and MI oocytes were as follows: 1 PN 0.8% vs 0.6%, P = 0.342; 3PN 2.6% vs 1.2%, P = 0.075; 0 PN 19.7% vs 45.2%, P &amp;lt; 0.001. Interestingly, the blastulation rate of zygotes with normal fertilization was very similar between both groups (46.4% vs 43.8%, P = 0.708), as well as the proportion of biopsied blastocysts on day 5 (74.2% vs 71.8%, P = 0.858), on day 6 (23.1% vs 25.6%, P = 0.946), and on day 7 (2.7% vs 2.6%, P = 0.594). The euploidy rate of the 298 biopsied MII-derived blastocyst was 36.2%, compared with 23.1% of the 39 MI-derived blastocysts biopsied (P = 0.077). Despite the limited sample size, it is noteworthy that three out of the four euploid blastocysts derived from MI oocytes, which were transferred in single embryo transfers, successfully led to pregnancy resulting in one healthy live birth, one ongoing pregnancy and one miscarriage at 9 weeks of pregnancy. Limitations, reasons for caution While this study provides valuable insights into the developmental competence of MI oocytes, it is not exempt from certain limitations, such as its retrospective nature, limited sample size and ethical considerations regarding the injection of putative immature oocytes. Wider implications of the findings MI oocytes showed lower fertilization rates compared to their MII siblings. However, once fertilized, MI-derived embryos had similar blastulation rates and a trend towards lower euploidy rates. Our study suggests that MI oocytes may result in successful pregnancies and live birth, but further research is needed to validate these findings. Trial registration number Not applicable

P-185 Shortening the time from ovulation trigger to insemination is effective in reducing multiple pronuclei formation in anti-centromere antibody-positive patients

Abstract Study question Does shortening the time from ovulation trigger to insemination reduce multiple pronuclei formation (MPF) and improve the outcome after fertilization? Summary answer Shortening the time from ovulation trigger to insemination increased the normal fertilization rate and increased the number of cycles in which good embryos were obtained. What is known already When anti-centromere antibody (ACA)-positive patients undergo IVF, low oocyte maturation rates and MPF may be observed, which tend to occur when ACA antibody titers are high. In this case, the patient tends to present with poor embryonic development. There is currently no effective treatment for ACA-positive patients. It has been reported that MPF is caused by chromosome dispersion due to significant abnormal spindle deformation. In such cases, it has been reported that the oocyte spindle is often not visualized by polarized light microscopy. The oocyte is affected by ACA during the meiosis, but the mechanism is not well understood. Study design, size, duration A retrospective study was conducted on 764 mature oocytes from 174 cycles in 45 patients who underwent oocyte retrieval and inseminated by intra-cytoplasmic sperm injection (ICSI) with consent from 2009 to 2022. Patients were limited to those who tested positive for ACA by blood antinuclear antibody testing and whose oocytes occurred MPF. Participants/materials, setting, methods Oocyte retrieval was performed 36 hours after ovulation trigger. After ICSI, embryos were cultured until day 3 or blastocyst. The group was divided into two groups: the time reduced from ovulation trigger to ICSI (S group, n = 95) and the normal protocol (N group, n = 79). The normal fertilization rate, MPF rate, the rate of good embryo on day 3, the percentage of IVF cycles with good embryo, and clinical pregnancy rate per transfer were compared. Main results and the role of chance The mean time from maturation trigger to insemination was shorter in the S group than in the N group (38.2h vs. 41.8h, P &amp;lt; 0.01). The normal fertilization rate was significantly increased in the S group compared to the N group (49.9 vs. 40.7, P &amp;lt; 0.01) and the MPF rate was significantly decreased (39.1 vs. 28.3, P &amp;lt; 0.01). The rate of transferable embryo on day 3 tended to increase in the S group compared to the N group (70.6 vs. 61.9, P = 0.09). In addition, the percentage of IVF cycles with good embryo was significantly increased in the S group compared to the N group (57.4 vs. 28.4, P &amp;lt; 0.01). There was no difference in pregnancy rate per embryo transfer. There was no difference in the age of the wife at the time of oocyte retrieval. Limitations, reasons for caution The data were conducted from a single facility. and the limited number of cases, resulting in a small sample size. The process of visualizing the oocyte spindle was not performed in this study because it is time consuming and increases the time from ovulation trigger to insemination. Wider implications of the findings Shortening the time from maturation trigger to insemination was decreased MPF, suggesting that the oocyte spindle is affected by ACA in a time-dependent manner. It is recommended that ICSI be performed as soon as possible after oocyte retrieval. Trial registration number not applicable

P-107 Sperm DNA fragmentation impacts embryonic developmental potential but is not associated with PGT-A outcomes

Abstract Study question Does sperm DNA Fragmentation Index (DFI) affect embryonic development and influence Preimplantation Genetic Testing for Aneuploidy (PGT-A) outcomes in IVF? Summary answer DFI adversely affects early embryonic development, diminishing blastocyst potential. Nonetheless, PGT-A outcomes are unaffected, underscoring the importance of selecting optimal sperm for enhanced IVF results. What is known already Several major factors are thought to cause DFI. The relationship between sperm DFI and IVF results remains debated. Study design, size, duration This study was conducted from January 2018 to March 2023, included 56 married couples undergoing IVF after performing a sperm DFI test. The DFI was assessed at the first semen examination and categorized into three groups: low (≤15%), moderate (15-30%), and high (≥30%). Participants/materials, setting, methods Intracytoplasmic sperm injection (ICSI) was performed on 1006 mature oocytes, followed by blastocyst culture. A total of 278 blastocysts meeting freezing criteria were vitrified. Trophectoderm biopsies from 216 blastocysts underwent Next-Generation Sequencing for aneuploidy analysis. Statistical analysis utilized the Cochran-Armitage test to compare outcomes among the DFI groups. Main results and the role of chance The average age of the husbands and wives who underwent IVF after the DFI examination was 42.3±4.2 and 41.4±2.4 years old. Normal fertility rates after ICSI in low, moderate, and high DFI were 85.5%, 84.5%, and 83.1%, with no significant difference. Similarly, the blastocyst freezing rates were 35.4%, 26.3%, and 29.6%, and tended to decrease as the DFI value increased (p &amp;lt; 0.05). However, the euploid rates for PGT-A were 22.4%, 16.3%, and 18.2%. There were no statistically significant differences between the DFI groups in terms of euploidy. Limitations, reasons for caution Cases declining ICSI due to patient preference or historical medical reasons were excluded from this study. Wider implications of the findings This suggests that sperm DNA imperfections have a negative impact on the developmental process during the early stages of division, but during late stages of division, the ability to repair DNA works to differentiate into blastocysts with normal developmental potential. Trial registration number not applicable

P-103 Sperm selection for ICSI by rheotaxis and thermotaxis in In-Situ handmade microfluidics of fluidic walls: preliminary results of a Randomized Controlled Trial

Abstract Study question Does sperm selection by rheotaxis and thermotaxis in In-Situ handmade microfluidics of fluidic walls optimize ICSI outcomes and workflow? Summary answer In-Situ handmade rheo/thermotaxis microfluidics of fluidic walls is as effective and more time-efficient than centrifugation density gradients to support pre-implantation embryonic development to blastocyst. What is known already Microfluidics technologies stand as the latest sperm selection methodology for ICSI. To outline their application in ART we conceived a lab-on-a-chip approach for sperm selection by In-Situ handmade microfluidics of fluidic walls (isM) in the same ICSI plate which integrates two operative zones: one to allow seminal plasma dispersion and sperm thermotaxis and another one to allow sperm rheotaxis. Our microfluidics system can be prepared with any media suitable for ICSI, disregarding centrifugation, washing, plasticware and other cells. The isM protocol proved non-inferior to centrifugation density gradients (CDG) to support embryonic development and implantation in a previous sibling-oocyte study. Study design, size, duration The present RCT was approved by our ethical committee for research with medical products (CEIm approval number 2023/394). This preliminary analysis aims to detect deviations in the pre-defined primary outcomes: Normal Fertilization Rate, Day 5/6 usable blastocyst rate per MII, and per zygote. This analysis includes data from 21 couples undergoing an ICSI treatment in our center during December 2023-January 2024. Only patients without clinical or analytical suspicion of sperm DNA fragmentation were included. Participants/materials, setting, methods Patients were randomly allocated to isM (n 9) or to CGD (n 12) groups. All participants signed an informed consent. Sperm samples were processed directly after liquefaction. For CGD, samples were prepared in 80-40% PureSperm® layers by isopycnic centrifugation followed by washing and dilution steps. The isM system was prepared by displaying three microdroplets of different volumes following a L-shape interconnection and allowing a temperature gradient of 2ºC between the first and the second microdroplet. Main results and the role of chance Sperm characteristics showed the following total mean values for the isM and CGD groups, respectively: concentration (58,2±38,0 × 106/ml and 52,7±29,1 × 106/ml) and progressive sperm concentration (16,6±17,0 × 106/ml and 16,8±13,4 × 106/ml). Women average age was 36,7±2,9 isM and 35,8±2,5 CDG. This analysis includes a total of 168 MII oocytes (n = 69 isM and n = 99 CDG), with a mean number of MII oocytes per patient of 7,4±2,1 isM and 8,3±1,9 CDG. Comparisons between the two groups showed that isM protocol optimizes ICSI outcomes by means of Normal Fertilization Rate: 88,4% vs 76,8%; p &amp;lt; 0,05, Day 5 A/B grade blastocyst rate: 36,1 vs 31,6%; ns, Day 5/6 usable blastocyst rate per MII: 58,0% vs 44,4%; ns and Day 5/6 usable blastocyst rate per diploid zygote: 65,6% vs 57,9%; ns. The isM protocol also optimizes the IVF workflow compared to CDG, as the time to obtain sperm for ICSI after liquefaction was 26 vs 77 minutes, respectively. Limitations, reasons for caution The study aims for a total size of 50 ICSI cycles to validate statistically our data at a power of 80%. The isM protocol depends on a free hand preparation. A ready-to-use surface would benefit the operability, albeit the use of templates and the training reduces inter and intra-operator variability. Wider implications of the findings The potential benefits of the isM protocol on fertilization could be significant for patients with a limited number of available oocytes for ICSI. According to the literature, microfluidics reduce the DNA fragmentation index in the selected sperm fraction. The isM method could be especially beneficial in patients with this pathology. Trial registration number NCT number assignment pending

O-018 Utilisation of second successive ejaculate as a way to overcome poor semen parameters for couples who wish to undergo conventional-IVF

Abstract Study question Can a second successive-ejaculate (SSE) be an option for patients with poor sperm parameters (PSPs) on ovum pick-up (OPU)-day who prefer to undergo conventional-IVF (c-IVF)? Summary answer SSE can be a valid option for patients with PSPs who do not wish to undergo ICSI as an assisted reproductive technology (ART) treatment. What is known already ART is applied when first-line treatment for infertility (timed intercourse or intrauterine insemination) is unsuccessful. The male partner is requested to follow ejaculatory abstinence, which according to the WHO guidelines is 2-7 days, while ESHRE recommends 3-4 days. Among these couples there are cases that require ICSI due to semen analysis with PSPs on the day of OPU. However, some couples are reluctant to undergo ICSI and prefer c-IVF as a treatment method. For those cases, we request a second semen sample on the same day (time interval: 1-4 h) and c-IVF is performed using the two ejaculates combined. Study design, size, duration This is a single-centre retrospective study employing 212 semen from male partners with PSPs in the first semen collection on the OPU-day and subsequently, an SSE was requested in order to perform c-IVF according to patients’ wishes. The data were collected from May 2006 to October 2023 in our clinic (SSE-group). The Control group included 271 semen samples from men where c-IVF was conducted using one ejaculate from January to December 2021 in our clinic. Participants/materials, setting, methods The sperm parameters were analysed according to the WHO manual for the examination and processing of human semen and compared between the first and second ejaculates in the SSE-group. Fertilisation rate (FR), percentage of clinically utilised embryos and clinical pregnancy rate were compared between the SSE-group and the Control group. The SSE-group had 254 ART-cycles and the mean maternal age was 36.8±4.9y.o. The Control group included 432 ART-cycles and the mean maternal age was 38.1±5.2y.o. Main results and the role of chance In SSE-group the semen volume of the first ejaculate was significantly higher compared to the second ejaculate (P&amp;lt; 0.0001). The sperm concentration and sperm motility were significantly higher in the second ejaculate compared to the first one (P&amp;lt;0.0001). In the same fashion, the total-motile sperm-count was significantly higher in the second ejaculate compared to the first (P=0.01). All the semen parameters in the Control group were significantly higher compared to both first and second ejaculates from the SSE-group (P&amp;lt;0.00001). The normal FR was significantly higher in the Control group compared to the SSE–group (P&amp;lt;0.01). On the other hand, there was no significant difference in the percentage of embryos that could be utilized (cryopreserved + freshly-transferred) between the two groups (P=0.218). Pregnancy rate for fresh embryo transfers (ETs) was significantly higher in the SSE-group compared to the Control (P=0.005), while there was no significant difference between the two groups for pregnancy rate for frozen/thawed ETs (P=0.728). The main strength of our study is that the utilisation of the SSE combined with the first ejaculate did not decrease the pregnancy rate for the couples with PSPs that wished to follow c-IVF as their treatment and it could actually provide an effective option for them. Limitations, reasons for caution SSE may improve the sperm parameters and give the opportunity to infertile couples to undergo c-IVF according to their wish. However, since the FR was lower in the SSE-group, it is of paramount importance to provide sufficient explanation to those couples and obtain their consent when requesting an SSE. Wider implications of the findings Overall, SSE had positive effect to a large number of parameters evaluated in this study. Additionally, the rate of utilised embryos was similar between the two groups. Two successive-ejaculates combined can be employed without any negative impact for couples with PSPs on the OPU-day that are reluctant to undergo ICSI. Trial registration number not applicable

P-028 Pregnancy and perinatal outcomes in male patients with temporary ejaculation failure undergoing assisted reproductive technology

Abstract Study question When temporary ejaculation failure occurs on the oocyte retrieval day, choose surgical sperm retrieval or oocyte cryopreservation? Summary answer TESA-ICSI and oocyte cryopreservation are both relatively efficient and safe way. Embryos have better developmental potential through TESA-ICSI. What is known already Nearly 9% of men among couples who use assisted reproductive technology have temporary ejaculation failure (TEF) on the oocyte retrieval day. If they fail to provide semen sample, the couples may face three choices (1. surgical sperm retrieval; 2. oocyte cryopreservation; 3. canceling the oocyte retrieval). The aim of the study was to compare the impact of testicular sperm aspiration (TESA) or oocyte cryopreservation on the clinical outcomes of intracytoplasmic sperm injection (ICSI) patients with TEF. Study design, size, duration The retrospective study included 345 male patients with TEF in our University-affiliated IVF center from 2015 to 2021. Participants/materials, setting, methods Three hundred and forty-five male patients were divided into two groups. Two hundred patients chose TESA-ICSI (the TESA group), while another 145 patients chose oocyte cryopreservation, waiting for the man’s next successful sperm ejaculation and subsequent ICSI (the frozen oocyte group). We investigated the embryo development, pregnancy and perinatal outcome, and included the data from fresh embryo transfer (ET) cycle. Herein, all patients in the frozen oocyte group received hormone replacement and fresh ET. Main results and the role of chance Firstly, there were no significant difference in the female BMI, FSH, antral follicle count (AFC), AMH, and male semen quality between the two groups. For embryo development, both two groups have the similar normal fertilization rate (71.5% vs 69.7%, P = 0.242). However, the frozen oocyte group has lower rate of Day3 high quality embryo (31.1% vs 49.3%, P &amp;lt; 0.001), available embryo rate (49.8% vs 55.8%, P = 0.043), and available blastocyst rate (18.6% vs 33.7%, P &amp;lt; 0.001). For pregnancy outcome, the clinical pregnancy rate (44.2% vs 50.0%), pregnancy loss rate (4.8% vs 2.9%) and live birth rate (38.5% vs 46.1%) were similar. And there were no significant differences in the rates of Gestational diabetes mellitus, hypertensive disorders of pregnancy, placenta previa and fetal malformation between the two groups. Limitations, reasons for caution This study only included data from fresh embryo cycles, not yet from thawed embryo cycles. Although we presented complete embryo development data for each couple, there may be some bias in pregnancy and perinatal outcomes due to the lack of data from thawed embryo cycles. Wider implications of the findings The present study suggested that TESA-ICSI and oocyte cryopreservation are both relatively efficient and safe way to help the patients encountering TEF. Due to the embryos with better developmental potential through TESA-ICSI, choosing the surgical sperm retrieval might have more have more advantages, and this has to be further verified. Trial registration number REC No. (2023(S142))

Novel insights into necrozoospermia from a single-center study: reference ranges, possible etiology, and impact on male fertility.

Necrozoospermia is a poorly documented condition with a low incidence, and its definition and clinical significance are unclear. Herein, we provide a reference range for necrozoospermia and discuss its possible etiology and impact on male fertility and assisted reproductive outcomes. We extracted relevant information from 650 Chinese male partners of infertile couples and statistically analyzed sperm vitality. Necrozoospermia was present in 3.4% (22/650) of our study population, and the lower cut-off value for sperm vitality was 75.3%. We compared two methods for assessing sperm vitality (eosin-nigrosin head staining and hypo-osmotic swelling test [HOST]), for which the percentage in the eosin-nigrosin group (mean ± standard deviation [s.d.]: 77.5% ± 10.5%) was significantly higher than that in the HOST group (mean ± s.d.: 58.1% ± 6.7% [5-10 min after incubation] and 55.6% ± 8.2% [25-30 min after incubation]; both P < 0.001). The incidence of necrozoospermia increased with age (odds ratio [OR] = 1.116, 95% confidence interval [CI]: 1.048-1.189, P = 0.001), while the percentage of normal sperm morphology and DNA fragmentation index (DFI) were significantly associated with necrozoospermia, with ORs of 0.691 (95% CI: 0.511-0.935, P = 0.017) and 1.281 (95% CI: 1.180-1.390, P < 0.001), respectively. In the following 6 months, we recruited 166 patients in the nonnecrozoospermia group and 87 patients in the necrozoospermia group to compare intracytoplasmic sperm injection (ICSI) and pregnancy outcomes between the two groups. The necrozoospermia group had a significantly lower normal fertilization rate (74.7% vs 78.2%, P = 0.041; OR = 0.822; 95% CI: 0.682-0.992) than that in the nonnecrozoospermia group. This study presents substantial information on necrozoospermia to establish comprehensive and applicable reference values for sperm vitality for spontaneous conception and artificially assisted reproductive management.

Effect of baseline prolactin levels on the pregnancy outcomes after fresh embryo transfer.

There is clinical disagreement on whether to treat hyperprolactinemia with medication before embryo transfer. The aim of this study is to identify the impact of basal prolactin (PRL) levels on pregnancy outcomes in fresh embryo transfer cycles. This retrospective study involved 2,648 women who underwent basal PRL level testing and fresh embryo transfer between January 2015 and December 2020 at our Hospital's Department of Assisted Reproduction. Basal PRL levels can be classified into three categories:<30 (n=2339), 30‑60 (n=255), and ≥60 (n=54) µg/l. Pregnancy outcome was defined as the primary outcome measure, and the live birth rate was defined as the second outcome measure. Subsequently, univariate and multivariable logistic regression analysis was used to reveal the association between basal PRL levels and pregnancy outcomes after considering several potential confounding factors. Elevated basal PRL levels were found not a risk factor for pregnancy outcomes in patients receiving good-quality embryo transfer (p>.05). For pregnancy or not, female age (OR: 1.03; 95% CI: 1.01-1.05), embryos transferred (OR: 0.52; 95% CI: 0.41-0.65), and normal fertilization rate (OR: 0.68; 95% CI: 0.48-0.97) were found to be an independent risk factor. For ongoing pregnancy or not, female age (OR: 1.07; 95% CI: 1.03-1.11), embryos transferred (OR: 0.57; 95% CI: 0.37-0.88), and menstrual cycle (OR: 1.76; 95% CI: 1.22-2.54) were also independent risk factors. There is no adverse impact on pregnancy outcomes during embryo transfer cycles with good-quality embryos when PRL levels are elevated.

Diverse impacts of female chromosomal polymorphisms on assisted reproduction outcomes: a retrospective cohort study

BackgroundThe effects of female chromosomal polymorphisms (FCPs) on various aspects of reproductive health have been investigated, yet the findings are frequently inconsistent. This study aims to clarify the role of FCPs on the outcomes of in vitro fertilization (IVF) and intracytoplasmic sperm injection (ICSI).MethodsThis retrospective cohort study comprised 951 couples with FCPs and 10,788 couples with normal karyotypes who underwent IVF/ICSI treatment at Peking University Third Hospital between 2015 and 2021. The exposure was FCPs. The embryological outcomes and clinical outcomes were compared.ResultsThe FCPs, as a whole, compromised the oocyte maturation rate (76.0% vs. 78.8%, P = 0.008), while they did not adversely affect other IVF/ICSI outcomes. Further detailed analyses showed that every type of FCPs contributed to the lower oocyte maturation rate, particularly the rare FCPs (69.0% vs. 78.8%, P = 0.008). The female qh + was associated with a higher normal fertilization rate (63.0% vs. 59.2%, adjusted P = 0.022), a higher clinical pregnancy rate (37.0% vs. 30.7%, adjusted P = 0.048), and a higher live birth rate (27.0% vs.19.0%, adjusted P = 0.003) in couples undergoing IVF. Conversely, in couples undergoing ICSI, female qh + was found to be related to a lower normal fertilization rate (58.8% vs. 63.8%, P = 0.032), a comparable clinical pregnancy rate (25.7% vs. 30.9%, P = 0.289), and a comparable live birth rate (19.8% vs. 19.2%, P = 0.880) compared to the control group. Additionally, an increased risk of preterm birth was observed in women undergoing IVF with multiple polymorphisms (62.5% vs. 16.9%, adjusted P < 0.001) and in women undergoing ICSI with pstk+ (36.4% vs. 15.4%, P = 0.036).ConclusionsOur research unravels the diverse impacts of various FCPs on IVF/ICSI outcomes, highlighting the detrimental effects of FCPs on oocyte maturation and the risk of preterm birth.

Polyvinylpyrrolidone can prevent oolemma lysis caused by abnormal rupture of the plasma membrane in Piezo-ICSI.

The aim of this study was to investigate whether it is possible to prevent oolemma lysis after Piezo-assisted intracytoplasmic sperm injection (Piezo-ICSI) caused by abnormal membrane rupture. A total of 489 mature oocytes were obtained from 85 patients who underwent Piezo-ICSI in an infertility clinic. Inseminated oocytes were classified into the following two groups: normal rupture and abnormal rupture, and a portion of abnormally ruptured oocytes was randomly exposed to 7% PVP solution. Normal fertilization rate, degeneration rate, proportion of high-quality embryos on day 3, blastocyst formation, and morphologically high-quality blastocysts were compared. Abnormal rupture was defined as cytoplasmic membrane rupture before piezo pulse driving. Among the abnormal rupture groups, the normal fertilization and degeneration rates were compared between the high-stretched (ruptured at ≥ 50% during oocyte membrane stretching) and low-stretched (< 50% position) oocytes.The degeneration rate was significantly higher in abnormally ruptured oocytes than in normally ruptures oocytes (14.3% vs 1.3%, p < 0.001), and there was no significant difference in embryonic development after fertilization. PVP treatment immediately after oolemma rupture significantly decreased the degeneration rate (6.0% vs 19.7%, p = 0.031) and increased the normal fertilization rate. Similarly, there were no significant differences in the developmental potential. Furthermore, pregnancy outcome data showed no significant differences in pregnancy and live birth rates. The degeneration rate was reduced by treating low-stretched oocytes with PVP.Exposure to polyvinylpyrrolidone (PVP) immediately after abnormal membrane rupture represents an effective strategy to prevent oocyte degeneration. This is the first study to propose a strategy for the prevention of oocyte degeneration in Piezo-ICSI. These findings are expected to reduce the oocyte degeneration rate and increase normal fertilization rate as well as assist patients who can only acquire oocytes with weak plasma membranes.

Impacts of male chromosomal polymorphisms on semen quality and IVF/ICSI outcomes: A retrospective cohort study.

The study aims to elucidate the impacts of different types of male chromosomal polymorphisms (MCPs) on various outcomes of invitro fertilization (IVF) and intracytoplasmic sperm injection (ICSI) treatment. This retrospective cohort study included 1442 couples with normal karyotypes, 1442 couples with MCPs, 42 couples with male chromosomal rearrangements (MCRs), and 42 couples with MCRs combined with MCPs who underwent IVF/ICSI treatment at Peking University Third Hospital from 2015 to 2021. The semen quality, embryological outcomes, and clinical outcomes of different groups stratified by karyotypes were compared. For couples undergoing IVF, male inv(9) was associated with a significantly lower sperm viability rate (29.41% vs 34.49%, P = 0.030), a lower progressive motility rate (25.13% vs 30.50%, P = 0.013), and a lower normal fertilization rate (52.41% vs 59.84%, P = 0.014). Male 9qh + was related to a lower sperm viability rate (27.56% vs 34.49%, P = 0.028). No MCPs were observed to compromise clinical outcomes in couples undergoing IVF. For couples undergoing ICSI, no MCPs exhibited an association with poorer semen quality and embryological outcomes. However, Yqh + and DGpstk+ were found to be significantly correlated with an increased likelihood of preterm birth (23.3% vs 9.2%, P = 0.003; 20.0% vs 9.2%, P = 0.041, respectively). In couples with MCRs, the presence of MCPs significantly reduced the sperm viability rate (19.99% vs 30.97%, P = 0.017) and progressive motility rate (8.07% vs 27.85%, P = 0.018). Our study provides detailed evidence for the impacts of various MCPs on IVF/ICSI outcomes, reveals the complexity and heterogeneity of these impacts, and highlights the adverse effects of male inv(9).