Normal Fertilization Rate Research Articles

Are testicular sperms superior to ejaculated sperms in couples with previous ART failure due to high rate of fragmented embryos? A retrospective cohort study.

The aim was to clarify whether using testicular sperm reduces embryo fragmentation and improves cycle outcomes. Fragmented embryo was defined as an embryo in which fragments account for more than one third of the embryonic surface area. High rate of fragmented embryos was defined by a proportion of fragmented embryos higher than 50%. We recruited infertile couples who had undergone at least one ovarian stimulation cycle using ejaculated sperm but failed to conceive due to high rate of fragmented embryos in each previous cycle. After fully informed consent, the couples agreed to obtain testicular sperm by testicular puncture and use testicular sperm for intracytoplasmic sperm injection (ICSI). The normal fertilization rate, transferable embryo rate, fragmented embryo rate and cycle outcomes were compared between ejaculated sperm group (EJA-sperm group) and testicular sperm group (TESTI-sperm group). Twenty-two couples who agreed to participate in our study underwent 32 ICSI cycles with ejaculated spermatozoa and 23 ICSI cycles with testicular spermatozoa. Embryo transfers were cancelled in 8 ejaculated cycles and 4 testicular cycles because of no transferable embryos. There were no significant differences in age, normal fertilization rate and high-quality embryo rate between ejaculated and testicular groups. The transferable embryo rate and implantation rate in TESTI-sperm group were significantly higher than those in EJA-sperm group (36.9% vs. 22.0%, p < 0.01; 34.2% vs. 0%, p < 0.001). The fragmented embryo rate in TESTI-sperm group was significantly lower than that in EJA-sperm group (61.2% vs. 75.7%, p < 0.05). Our small retrospective cohort study suggests that using testicular sperm may be a recommended option for couples with previous ART failure because of high rate of fragmented embryos. Large samples, multicenter studies or randomized controlled trial (RCT) are needed to further confirm the superiority of testicular sperm.

The influence of male and female overweight/obesity on IVF outcomes: a cohort study based on registration in Western China

BackgroundOverweight/obesity can affect fertility, increase the risk of pregnancy complications, and affect the outcome of assisted reproductive technology (ART). However, due to confounding factors, the accuracy and uniformity of published findings on IVF outcomes have been disputed. This study aimed to assess the effects of both male and female body mass index (BMI), individually and in combination, on IVF outcomes.MethodsThis retrospective cohort study included 11,191 couples undergoing IVF. Per the Chinese BMI standard, the couples were divided into four groups: normal; female overweight/obesity; male overweight/obesity; and combined male and female overweight/obesity. The IVF outcomes of the four groups were compared and analysed.ResultsRegarding the 6569 first fresh IVF-ET cycles, compared with the normal weight group, the female overweight/obesity and combined male/female overweight/obesity groups had much lower numbers of available embryos and high-quality embryos (p < 0.05); additionally, the fertilization (p < 0.001) and normal fertilization rates (p < 0.001) were significantly decreased in the female overweight/obesity group. The combined male/female overweight/obesity group had significant reductions in the available embryo (p = 0.002), high-quality embryo (p = 0.010), fertilization (p = 0.001) and normal fertilization rates (p < 0.001); however, neither male or female overweight/obesity nor their combination significantly affected the clinical pregnancy rate (CPR), live birth rate (LBR) or abortion rate (p > 0.05).ConclusionOur findings support the notion that overweight/obesity does not influence pregnancy success; however, we found that overweight/obesity affects the fertilization rate and embryo number and that there are sex differences.

Cryptozoospermia: Should we use ejaculated sperm or surgically retrieved sperm for assisted reproductive technology?

In cryptozoospermic subjects, it may often may be difficult to secure motile sperm for assisted reproductive technology (ART). We examined the results of ART with frozen thawed ejaculated sperm in cryptozoospermic subjects and evaluated whether sperm retrieval surgery is necessary for such patients in our clinic. Between 2013 and 2021, we evaluated 197 cryptozoospermic patients. Age, endocrine panel at the time of the initial semen analysis, and anti-müllerian hormone levels at the time of the spouse's first egg retrieval were examined. Cryopreservation of ejaculated motile sperm collected essentially weekly over a 3-month period was carried out. ART data recorded was the number of egg retrieval cycles, normal fertilization rate, and clinical pregnancy rate. ART using frozen sperm as well as sperm ejaculated on the day of egg retrieval was possible in all cases. The normal fertilization rate was 70.4%, the clinical pregnancy rate per embryo transferred was achieved in 31.5% (870 cycles), and the live birth rate per case was 73.8%. Intracytoplasmic sperm injection (ICSI) was possible without sperm retrieval surgery in cryptozoospermia, resulting in 73.8% of live births per patient. Sperm identification, sperm processing, and ICSI technique are especially important in cryptozoospermia. Sperm retrieval surgery can be avoided in cryptozoospermic patients.

Association of paternal MTHFR polymorphisms (C677T) with clinical outcomes in ICSI treatment.

This study aims to investigate the association between paternal methylenetetrahydrofolate reductase (MTHFR) polymorphisms (C677T) and embryonic development, pregnancy, and neonatal outcomes in intracytoplasmic sperm injection (ICSI) treatment. A total of 191 infertile men undergoing ICSI treatment at the Reproductive and Genetic Hospital, The First Affiliated Hospital of USTC, were recruited between January 2020 and June 2021. The MTHFR C677T polymorphism genotyping was evaluated in these male patients, and they were stratified into three groups according to genotyping results: Control (CC), heterozygote mutated (CT), and mutated homozygote (TT). In addition, we conducted a comparative analysis of embryonic development, pregnancy, and neonatal outcomes among these three groups. The embryonic development (including normal fertilization rate (80.14% vs. 83.06% vs. 85.10%; p = 0.37), high-quality embryo rate (45.26% vs. 43.69% vs. 46.04%; p = 0.72), blastocyst formation rate (42.47% vs. 43.18% vs. 39.38%; p = 0.62), implantation rate (42.47% vs. 36.25% vs. 41.22%; p = 0.62), and clinical pregnancy rate (64.71% vs. 58.75% vs. 66.67%; p = 0.59) were not comparable among these three groups. Moreover, no significant difference was observed in terms of pregnancy outcomes (including miscarriage rate (24.24% vs. 12.77% vs. 22.5%; p = 0.35) and live birth rate (49.02% vs. 51.25% vs. 51.66%; p = 0.96)). Additionally, no marked difference was observed in terms of neonatal outcome (including, preterm delivery rate (24% vs. 14.63% vs. 9.67%; p = 0.35), birth height (p = 0.75), birth weight (p = 0.35), neonatal sex (p = 0.48), gestational age at delivery (p = 0.24), Apgar score (p = 0.34), and birth defects (0% vs. 2% vs. 9%; p = 0.23) among the study groups. The paternal MTHFR C677T polymorphism is not associated with embryo quality, pregnancy, or neonatal outcomes in ICSI treatment. Therefore, in our population, MTHFR polymorphisms do not provide helpful information in explaining ICSI failure.

Association between maternal MTHFR C677T/A1298C combination polymorphisms and IVF/ICSI outcomes: a retrospective cohort study.

What are the roles of maternal 5,10-methylenetetrahydrofolate reductase (MTHFR) C677T/A1298C combination polymorphisms on the embryological and clinical outcomes of IVF/ICSI? Our study reveals for the first time that the oocyte maturation potential gradually decreases with a reduction of maternal MTHFR activity determined by combined C677T/A1298C polymorphisms, while embryo quality was worse in women with intermediate MTHFR activity. Although many previous studies have explored the association between MTHFR polymorphisms and IVF/ICSI outcomes, the results remain contradictory due to inadequate samples, no adjustment for potential confounders and/or the study of C677T and A1298C separately. Few studies have systematically investigated the exact role of MTHFR activity determined by combined C677T/A1298C polymorphisms on the embryological and clinical outcomes of IVF/ICSI. This is a retrospective cohort study investigating 1160 women who were referred for MTHFR genotyping and IVF/ICSI treatment at Peking University Third Hospital from May 2017 to May 2020. Women who were referred for MTHFR genotyping and their first IVF/ICSI treatment at our hospital were included and those undergoing preimplantation genetic testing cycles were excluded. The included women were divided into different cohorts according to their C677T, A1298C and combined C677T/A1298C genotypes. The embryological outcomes, including oocytes retrieved, metaphase II oocytes, oocyte maturation rate, normal fertilization rate and transplantable embryo rate, were evaluated by generalized linear regression models. The clinical outcomes, including biochemical pregnancy rate, clinical pregnancy rate and live birth rate, were evaluated by log-binomial regression models. All outcomes were adjusted for potential confounders. Women with the combined 677TT/1298AA genotype (hereafter abbreviated as TT/AA, as with other combined genotypes), whose enzyme activity was the lowest, had a lower oocyte maturation rate compared with those with the wild-type genotype (P = 0.007). Moreover, the oocyte maturation rate decreased linearly with the decline in MTHFR enzyme activity determined by combined C677T/A1298C genotypes (P-trend = 0.001). The combined CC/AC, CC/CC&CT/AA and CT/AC genotypes with intermediate enzyme activity were associated with a lower transplantable embryo rate (P = 0.013, 0.030 and 0.039, respectively). The differences in clinical outcomes between women with wild-type genotype and combined C677T/A1298C variant genotypes were not significant. Our study population had comparable embryological outcomes but worse clinical outcomes than other women undergoing IVF/ICSI treatment at our hospital. Therefore, the results related to the clinical outcomes should be generalized with caution. In addition, we did not detect the folate concentration of each patient during pregnancy. However, this might not have much influence on our results because almost all of our study participants took sufficient folic acid around pregnancy. We provide a holistic view of the effect of MTHFR C677T and A1298C polymorphisms on the IVF/ICSI outcomes, which can contribute to providing reasonable folic acid supplementation suggestions for women with different MTHFR genotypes, especially for those with a low oocyte maturation rate and/or low embryo quality. This work was funded by the National Natural Science Foundation of China (31871447, and 82101677), the National Key Research and Development Program (2019YFA0801400) and the Natural Science Foundation of Beijing Municipality (7202226). The authors declare that they have no competing interests. N/A.

Maternal supplementation of nano-selenium in a plant-based diet improves antioxidant competence of female Arabian yellowfin sea bream (Acanthopagrus arabicus) breeders and their progeny

Antioxidants such as selenium (Se) play vital roles in reproduction success and larval development in fish. A three-month feeding experiment was conducted to examine the impact of enriching a plant-based diet (60% of fishmeal was substituted with a blend of plant ingredients) with nano-selenium (nano-Se) on antioxidant metabolism in female brooders and the progeny of Arabian yellowfin sea bream (Acanthopagrus arabicus). At this point, the plant-based diet was supplemented with graded levels of nano-Se at 0, 0.5, 1, 2, and 4 mg/Kg diet. Moreover, a fishmeal-based diet served as a positive control (FMD-Control). Broodfish were randomly distributed into eighteen 10 m3 rectangular concrete tanks (8 males and 8 females in each tank). Each experimental diet was subjected to three replications. Selenium retention increased in the serum, liver, ovary, eggs, and three-day larvae with increasing dietary nano-Se levels (P < 0.05). Supplementing the plant-based diet with 2–4 mg nano-Se/Kg significantly enhanced normal embryogenesis, fertilization, hatching, and larval survival rates. Generally, fish fed on plant-based diets with lower nano-Se supplementation (0–0.5 mg/Kg) had higher catalase and superoxide dismutase activities in the liver, ovaries, eggs, and larvae compared to the other groups. Glutathione peroxidase activity and total antioxidant capacity markedly increased, whereas lipid peroxidation decreased in the liver, ovary, serum, eggs, and progeny of broodfish fed with nano-Se supplemented diets (P < 0.05). In conclusion, supplementation of 2–4 mg nano-Se /Kg in a plan-based feed is recommended for the improvement of antioxidant defense in female A. arabicus brooders and their progeny.

Hsa-miR-320a-3p and hsa-miR-483-5p levels in human granulosa cells: promising bio-markers of live birth after IVF/ICSI

BackgroundMicroRNAs (miRNAs) are considered potential biomarkers for various diseases. This study investigated whether hsa-miR-320a-3p and hsa-miR-483-5p levels in human ovarian granulosa cells derived from follicular fluids are associated with embryo developmental competence.MethodsWe collected 195 granulosa cells samples and analyzed the treatment outcomes in patients undergoing in vitro fertilization (n = 147) or intracytoplasmic sperm injection (n = 48) cycles. The hsa-miR-320a-3p and hsa-miR-483-5p levels in granulosa cells were measured using quantitative reverse transcription-polymerase chain reaction.ResultsPatients were subdivided into four groups according to the granulosa cells hsa-miR-320a-3p and hsa-miR-483-5p levels quartiles (Q1–Q4). Embryo developmental competence was compared using the chi-square test. Patients in Q3 were less likely to achieve a normal fertilization rate for in vitro fertilization and blastocyst formation than those in Q1 as they expressed high levels of hsa-miR-320a-3p and hsa-miR-483-5p (P < 0.05). Patients in Q3 and Q4 were less likely to achieve a good-quality embryo as they expressed high levels of hsa-miR-483-5p and hsa-miR-320a-3p (P < 0.05). The hsa-miR-320a-3p and hsa-miR-483-5p levels were not associated with clinical pregnancy. However, multiple regression analysis indicated that in Q3 and Q4 intervals had experienced a decreased chance of live birth due to high expression levels of hsa-miR-320a-3p and hsa-miR-483-5p levels. The relative hsa-miR-320a-3p expression levels in granulosa cells were weakly and positively correlated with the patient age (P = 0.0033). Moreover, both the basal follicle stimulating hormone (P = 0.0003) and ovarian stimulation protocols (P = 0.006 and P = 0.004) significantly and positively affected hsa-miR-320a-3p levels. The days of stimulation was negatively correlated with the relative hsa-miR-320a-3p expression level (P = 0.047).ConclusionsThe hsa-miR-320a-3p and hsa-miR-483-5p levels in human granulosa cells negatively correlated with the good-quality embryo rate and live birth, indicating that hsa-miR-320a-3p and hsa-miR-483-5p can be used as potential negative indicators to predict good-quality embryos and live births.

Impact of paternal age on IVF and pregnancy outcomes with only normal sperm parameters

ObjectivePrevious reports on advanced paternal age effects on assisted reproductive technology (ART) vary considerably and those on frozen–thawed embryo transfer (FET) are rare. We investigated whether paternal age affects in vitro fertilisation (IVF) and FET pregnancy outcomes. Materials and methods1657 IVF cycles performed from January 2014 to May 2018 were retrospectively investigated excluding cases of poor semen parameters. Paternal and maternal ages were categorised into groups, namely, <35, 35–39 and ≥ 40 years, to compare normal fertilisation (2 PN (pronuclei)) and high-quality blastocyst rates. Furthermore, 741 FET cycles were investigated and pregnancy, live birth and miscarriage rates were compared. ResultsFor the maternal age group (35–39), the 2 PN rate was significantly higher with paternal age group of <35 than groups of 35–39 and ≥ 40 (median%, <35 vs. 35–39 vs. ≥40 = 100.0 vs. 71.4 vs. 77.7; P = 0.005). The miscarriage rate was significantly higher with paternal age group of ≥40 than that of <35 and 35–39 when maternal age was <35 (median %, <35 vs. 35–39 vs. ≥40 = 13.1 vs. 7.8 vs. 33.3; P = 0.038). ConclusionOur findings show that when maternal age was <35, advanced paternal age reduces the normal fertilisation rate and increases the FET miscarriage rate when maternal age was 35–39.

A novel solution for freezing individual spermatozoa using a right angular cryopiece embedded in a grooved petri dish.

Herein, we introduced a novel individual sperm freezing device named SpermCD, which consists of a right angular cryopiece (RA-Cryopiece, or "C") and a grooved petri dish ("D"). SpermCD allows embryologists to transfer sperm and perform ICSI on the same focal plane. Thirty-five patients underwent single sperm cryopreservation using SpermCD, including four patients with non-obstructive azoospermia (NOA), 14 patients with virtual azoospermia and 17 patients with cryptozoospermia. One hundred and twenty-five cryopreserved spermatozoa from nine patients were thawed on the day of the oocyte retrieval and 121 spermatozoa were found, with a sperm recovery rate of 97.1± 4.6%. Sixty-five MII oocytes from their spouse were injected with thawed sperm. Normal fertilization and high-quality embryo rates were 68.0% ± 33.2% and 24.4% ± 22.2%. Nineteen transplantable embryos were formed after fertilization with frozen sperm, eight of which were transplanted in five couples, resulting in four successful deliveries. SpermCD is a simple and practical individual sperm freezing device.

Group or individual housing does not reduce socio-sexual and reproductive responses in anestrous goats during the first contact with the photo-stimulated buck

Herein, we examined the effect of the type of housing (individual vs group housing) of anestrus female goats on plasma cortisol concentrations and socio-sexual behaviors during first contact with photo-stimulated male goats and determined the impact on sexual and reproductive responses after 15 d of contact with photo-stimulated males. Therefore, after weaning, 10 females each were individually and group-housed, respectively. Ten bucks were rendered sexually active by exposure to artificially long days (16 h of light and 8 h of darkness per day) for 2.5 mo. During the naturally increasing photoperiod, 15-mo-old females were exposed and maintained with males. On day 0, during the first contact with males, individually- and group-housed females exhibited similar plasma cortisol concentrations (22.6 vs 27.4 ng/mL, respectively). Likewise, socio-sexual behaviors did not differ between examined groups on day 0. Moreover, the interval from first contact with males to the first estrus, short and normal ovulatory cycles, ovulation, fertility, and prolificacy rates were similar between differently housed females. Furthermore, mounting attempts did not differ between males in contact with either female group. In conclusion, individually- and group-housed anestrus females displayed elevated and similar plasma cortisol concentrations during first contact with photo-stimulated males and similar socio-sexual behaviors and reproductive responses when exposed to photo-stimulated males.

Correlation of DNA fragments with routine semen parameters and lifestyle and their impact on assisted reproductive outcomes

ObjectiveThis study investigated the correlation between sperm DNA integrity and routine semen evaluation parameters in male infertile patients, the influencing factors, and the impact of the DNA fragmentation index (DFI) on embryo quality and clinical outcomes in in vitro fertilization and embryo transfer (IVF-ET). MethodsSperm DFI and semen routine parameters of 6160 infertile men admitted between June 2017 and June 2018 were analyzed. Patients were divided into three groups according to their DFI: low-DFI (DFI<15%), medium-DFI (15%<DFI≤30%), and high-DFI (DFI>30%). The correlations of DFI with patients’ age, sperm concentration, sperm percentage of forward movement and sperm percentage of normal shape were analyzed. The clinical data of 5040 infertile couples who received IVF treatment between June 2016 and 2021 and had embryos transferred in a fresh cycle were reviewed. The fertilization rate, cleavage rate, blastocyst rate, and pregnancy rate in different DFI groups were compared. ResultsSemen evaluation parameters (concentration, spermatozoa with progressive motility, and the normal morphology rate), the high-quality embryo rate, blastocyst development rate, and pregnancy rate in the high-DFI group were significantly lower than those in the other two groups. The correlation analysis revealed that sperm DFI was negatively correlated with semen concentration, sperm motility, and normal sperm morphology and positively correlated with the man's age, BMI, and unhealthy habits (smoking and drinking). There was no significant difference in the number of mature eggs and normal fertilization rate among groups. ConclusionA strong correlation exists between sperm DFI and semen evaluation parameters. Smoking, drinking, and other unhealthy habits lead to an increase in DFI, reducing the high-quality embryo rate and blastocyst development rate and affecting pregnancy outcomes.

Studying the IVF Laboratory Performance Indicators the Vienna Consensus 2017 for the High Institute According to Diagnosis and Assisted Reproductive for Infertility Technologies, Al-Nahrain University, Iraq

Background: Performance indicators are used to assess patient safety, efficacy, equity, patient-centeredness, punctuality, and efficiency. The benchmark values for each Key Performance Indicator are aspirational values, and the minimum performance level values are the number of fertilized oocytes on Day 1 and the Normal Fertilization Rate, respectively (presence of 2Pro Nucleus and 2Polar Body measured at 17 h post-injection) as a Failed fertilization rate is calculated as the proportion of IVF cycles (excluding cycles with intracytoplasmic injection) on Day 1 (17 hours after insemination) with no signs of pregnancy.&#x0D; During fertilization (i.e., 0 oocytes with 2Pro Nucleus). The percentage of zygotes on Day 2 (44 hours after insemination) is known as the cleavage rate, and it can suggest an issue with sperm quality (sperm function, oocyte activation, and gamete receptors), sperm processing, or the quantity of spermatozoa used for insemination. which cleaves to create embryos. The percentage of cleaved embryos per successfully fertilized egg that are at the 4-cell stage on Day 2 (44 hours post-insemination) or at the 8-cell stage on Day 3 (68 hours post-insemination) is known as the embryo development rate. This evaluates the viability and quality of the embryos as well as the capacity of the culture system to promote cleavage at the necessary stages. The critical factor is the proportion of blastocysts observed at 116 hours after insemination as a function of the number of correctly fertilized oocytes. Measures of performance blastocyst development rate. The viability of the embryo as well as the culture system's capacity to support blastocyst formation from fertilized oocytes (i.e., the formation of an intracellular mass and a blastocoele cavity) are both determined by this factor. It should be noted that this phrase only considers blastocyst formation and not blastocyst stage or quality. The damage rate is the proportion of oocytes that are injured or have deteriorated by the time of fertilization assessment on Day 1 as a result of the intracytoplasmic injection. The percentage of biopsied and tubed/fixed samples where DNA is found represents the success rate of the biopsy. It serves as a gauge of the embryologists' ability to transfer biopsied samples to test tubes, as shown by successful DNA amplification. The number of gestational sacs divided by the total number of transplanted embryos is how one calculates the implantation rate, which is dependent on the cleavage stage. By dividing the number of gestational sacs by the total number of transplanted blastocysts, the implantation KPI (blastocyst stage) is calculated.

Clinical efficacy analysis of oocyte cryopreservation: A propensity score matched study.

To investigate the effectiveness of oocyte thawing cycles in the clinical application of assisted reproductive technology (ART). The clinical data of 78 cases who underwent oocyte thawing cycles in our center were retrospectively analyzed. All patients in this study received oocyte cryopreservation for the husband reason. According to patient age at egg freezing, patients were divided into three observation groups (Group A, <30 years old; Group B, 30-34 years old; Group C, ≥35 years old), and the control groups were selected by propensity score matching with fresh cycles. The clinical outcomes of each group were compared, and the clinical efficacy of oocyte thawing cycles was analyzed. Clinical pregnancy outcomes of oocyte thawing cycles were not significantly different from that of fresh oocytes, but vitrification affected the number of two pronuclei zygotes developing to cleavage stage and the number of high-quality embryos, and the normal fertilization rate after thawing. The cycle cumulative live birth rate in Group C was significantly lower than those in Groups A and B. The live birth rates per egg of Groups A, B, C were 5.03%, 5.61%, and 3.57%, respectively, and the numbers of eggs per live birth were 13.72, 14.43, and 21.0, respectively. The overall clinical outcomes of oocyte vitrification were similar to that of fresh oocytes, but the cleavage rate and embryo quality of frozen oocytes were slightly reduced. Freezing of oocytes in women over 35 years of age affects the clinical efficacy of ART.

Rapamycin improves the developmental competence of human oocytes by alleviating DNA damage during IVM.

Can rapamycin improve the developmental competence of human oocytes during the IVM process? Rapamycin at 10 nM could markedly improve the developmental competence of human oocytes undergoing IVM. Embryos derived from oocytes that mature in vitro have lower developmental competence than sibling embryos derived from oocytes matured in vivo. Rapamycin was shown to effectively improve IVM outcomes in mammalian oocytes; however, its effects on IVM of human oocytes have not been investigated. In 2021, donated immature oocytes (n = 202) from 80 infertile couples receiving ICSI were included in a control group, and 156 oocytes from 72 couples were included in a rapamycin group. The oocytes underwent IVM with 10 nM rapamycin or without (control) rapamycin, followed by insemination by ICSI and embryo culture. The germinal vesicle breakdown (GVBD), maturation, normal fertilization, high-quality embryo (HQE) and blastocyst formation rates were calculated to evaluate the developmental competence of IVM oocytes, and fluorescence staining was used to assess DNA damage levels of oocytes in both groups. Whole-genome amplification and DNA sequencing were performed to analyze chromosome euploidy in embryos derived from the rapamycin group. The baseline characteristics of patients who donated oocytes for the two experimental groups were similar. In the control group, GVBD happened in 135 (66.8%) oocytes, and the maturation rate reached 52.5% at 24 h and 63.4% at 48 h. In the rapamycin group, 143 (91.7%) oocytes underwent GVBD, and the maturation rate reached 60.3% at 24 h and 82.7% at 48 h. Following ICSI, more HQEs were obtained in the rapamycin group versus control (34.2% versus 22.1%, respectively, P = 0.040), although with comparable fertilization rates in the two groups. In addition, the levels of histone γH2AX in oocytes cultured with 10 nM rapamycin were markedly decreased, compared with those in the control group (0.3 ± 0.0 versus 0.6 ± 0.1, respectively, P = 0.048). Embryos with normal karyotype could be obtained from oocytes cultured with rapamycin. Our preliminary results indicated that the addition of rapamycin during human oocyte IVM did not cause extra aneuploidy. However, this safety evaluation of rapamycin treatment was based on limited samples and more data are needed before possible application in the clinic. In the current study, 10 nM rapamycin was applied in the IVM process of human oocytes for the first time and showed positive effects, providing new insights for potentially improving IVM outcomes in the clinic. There were subtle differences between the results presented here on human oocytes and our previous studies on mouse oocytes, indicating the necessity of more research on human samples. This work was supported by the research grants from National Key Research and Development Project (2018YFC1002103) and Health Commission of Hubei Province scientific research project (WJ2021M110). All authors declared no conflicts of interest. N/A.

Oocytes With Smooth Endoplasmic Reticulum Aggregates May Not Impact Blastocyst Euploidy Rate.

ObjectiveTo investigate whether the euploidy rate of blastocysts derived from smooth endoplasmic reticulum aggregates (SERa) positive cycles and oocytes are impacted.DesignRetrospective cohort study.Method(s)A total of 601 preimplantation genetic testing (PGT) cycles with at least one oocyte retrieved in our center between April 2017 and May 2021 were initially included in the study. Women>35 years and PGT cycles with chromosomal structural rearrangements (PGT-SR) were excluded. Embryological and blastocyst ploidy outcomes were compared among SERa+ oocyte, sibling SERa- oocytes and oocytes in SERa- cycles.ResultsNo significant difference was observed among the SERa+ oocyte group, sibling SERa- oocyte group, and SERa- cycle group in the normal fertilization rate (82.1% vs. 77.8% vs. 83.1%, respectively, P=0.061), blastocyst formation rate (71.0% vs. 72.5% vs. 68.4%, respectively, P=0.393), good quality blastocyst formation rate (46.4% vs. 48.3% vs. 42.6%, respectively, P=0.198). No significant difference was observed in the euploidy rate (50.0% vs. 62.5% vs. 63.3%, respectively, P=0.324), mosaic rate (12.5% vs. 9.7% vs. 13.4%, respectively, P=0.506), and aneuploidy rate (37.5% vs. 27.8% vs. 23.2%, respectively, P=0.137) among the three groups.ConclusionOur results suggest that the euploidy rate of blastocysts derived from SERa+ cycles and oocytes may not be impacted.

Reproductive outcomes of dual trigger with combination GnRH agonist and hCG versus trigger with hCG alone in women undergoing IVF/ICSI cycles: a retrospective cohort study with propensity score matching

BackgroundDespite a large number of studies on the selection of trigger drugs, it remains unclear whether the dual trigger with human chorionic gonadotropin (hCG) and gonadotropin-releasing hormone (GnRH) agonist, compared to the trigger with hCG alone, can improve the reproductive outcome of patients undergoing assisted reproductive technology. Therefore, this study aimed to compare the laboratory and clinical outcomes of dual trigger versus single trigger.MethodsIn this retrospective cohort study, we evaluated 520 in vitro fertilization/intracytoplasmic sperm injection (IVF/ICSI) cycles between July 2014 and September 2020 at the Reproductive and Genetic Center of Integrative Medicine, The Affiliated Hospital of Shandong University of Traditional Chinese Medicine. All patients underwent IVF/ICSI treatment with fresh embryo transfer using the GnRH antagonist protocol. We used propensity score matching to control for confounding variables and binary logistic regression analysis to determine the correlations between trigger methods and pregnancy outcomes. After propensity score matching, 57 cycles from each group were evaluated and compared for laboratory or clinical outcomes in this retrospective cohort study.ResultsThere was no significant difference in the number of oocytes retrieved, embryos available, top-quality embryos, or the rate of normal fertilization between the dual-trigger and single-trigger protocols, respectively. The incidence of ovarian hyperstimulation syndrome, implantation rate, biochemical pregnancy rate, clinical pregnancy rate, ectopic pregnancy rate, early miscarriage rate, and live birth rate were also similar between the two groups, while the miscarriage rate (37.0% vs. 12.5%, p = 0.045) was higher in the dual-trigger than the single-trigger group. Subsequent binary logistic regression analysis showed that age was a remarkably significant independent predictor of both clinical pregnancy rate (odds ratio = 0.90, 95% confidence interval: 0.84–0.97, p = 0.006) and live birth rate (odds ratio = 0.89, 95% confidence interval: 0.82–0.97, p = 0.005).ConclusionsTherefore, dual-trigger for final oocyte maturation might increase miscarriage rate, but in terms of the laboratory and other pregnancy outcomes such as clinical pregnancy rate, early miscarriage rate or live birth rate, there was no evidence to show that dual trigger was superior to an hCG-trigger alone for patients undergoing GnRH-antagonist cycles with fresh embryo transfer.Trial registrationRetrospectively registered.

Effect Analysis of Degranulated Cell in Early Fertilization on FET Outcome and Offspring Safety with Data Mining.

In vitro fertilization and embryo transfer is one type of assisted reproductive technology, although the technology is now more mature. Many factors, however, will have an impact on oocyte fertilization, embryo growth, pregnancy outcome, and child safety due to the journey from clinical to the laboratory. The influence of degranulated cells early in fertilization on frozen embryo transfer (FET) results is investigated in this study. This article analyzes 255 patients who underwent in vitro fertilization (IVF) and FET transplantation at the author's central unit from January 1, 2015, to June 30, 2021. Among them, IVF-assisted conception is the early degranulation of homologous oocyte fertilization. Correlation analysis is performed by observing the embryonic outcome of the early degranulation group and the overnight fertilization group and the clinical outcome after FET. Through data mining analysis, the results show that the polyfertilization rate and 0PN rate for the early degranulation group are significantly higher than the overnight fertilization group (9.87% vs. 8.24% and 3.14% vs. 1.69%). In terms of normal fertilization rate, there is no significant difference between D3 high-quality embryo rate and D5 high-quality blastocyst rate (64.07% vs. 65.15%, 27.5% vs. 26.5%, and 15.97% vs. 17.35%). There is no significant difference in the complete recovery rate of embryos after thawing (93.24% vs. 93.46%), and the implantation rate, clinical pregnancy rate, abortion rate, and live birth rate are not significantly different between the two groups after FET. The offspring outcomes of singletons do not differ significantly between the two groups; however, twins born early degranulate have much greater rates of ultralow birth weight and ultrapreterm children than twins born overnight fertilization (14.29% vs. 0). Therefore, it can be concluded that degranulation of cells early in fertilization is a desirable method to prevent fertilization disorders. However, under the premise of ensuring that no fertilization disorder occurs, it is not appropriate to degranulate all the oocytes of the patient at the early stage of fertilization.

Study on the Influencing Mechanism of Human Chorionic Gonadotropin (hCG) on Oocyte Maturation in Patients with Polycystic Ovary Syndrome.

The study was aimed at investigating the influence of human chorionic gonadotropin (hCG) hormone on oocyte maturation in the patients with polycystic ovary syndrome (PCOS). A total of 54 patients with PCOS who received in vitro maturation (IVM) treatment in the Cheeloo College of Medicine, Shandong University, were divided into two groups: one group who underwent hCG injections was the observation group (OG; n = 27) and other was the control group (CG; n = 27) with no hCG injection. The oocyte development and the expression of steroid hormone synthesis-related genes including gonadotropin-releasing hormone receptor (GnRHR), Conexin43, epidermal growth factor-related genes, luteinizing hormone/choriogonadotropin receptor (LHCGR), epiregulin (EREG), and vascular endothelial growth factor (VEGF) were examined. The human ovarian granulosa cell line (SVOG cells) and ovarian epithelial cell line (HOSEpiC cells) were employed to analyze the effect of hCG on the biological behaviour of cells. As a result, OG showed higher normal fertilization, cleavage, and high-qualified embryo rate than CG. Expression levels of GnRHR, Cx43, LHCGR, EREG, and VEGF were significantly elevated in granulosa cells in the OG group. Western blot revealed that phosphoinositide 3-kinase (PI3K), protein kinase B (AKT), and rapamycin (mTOR) proteins were decreased in granulosa cells under hCG intervention. A biological behaviour test indicated that the multiplication capacity of hCG-intervened SVOG and HOSEpiC was increased, while the apoptosis was decreased. In conclusion, hCG could accelerate follicular development and oocyte maturation by activating oocyte maturation genes in PCOS patients, which could significantly improve and popularize the application of IVM technology.

P-022 Relationship between sperm movement characteristics assessed by a sperm motility analysis system (SMAS) and normal fertilization rates in vitro

Abstract Study question Is there a relationship between movement characteristics of human sperm assessed by SMAS and normal fertilization rates in vitro? Summary answer The curvilinear path wobble of motile sperm before insemination is predictive of normal fertilization rates after in vitro fertilization (IVF). What is known already Assessment of semen parameters is very important in both the diagnosis of male infertility and the selection of methods for treatment with conventional-IVF (C-IVF) or intracytoplasmic sperm injection (ICSI). Several studies have reported that objectively movement characteristics of motile sperm assessed by SMAS are related to the functional capacity of human sperm. However, there are no established the objective indicators and criteria for sperm motility characteristics assessed by SMAS. In this study, we investigated the utility of the movement characteristics of motile sperm measured by SMAS as an indicator of normal fertilization rates. Study design, size, duration Retrospective experimental trial conducted in a single IVF center between January 2018 and February 2020. Sperm movement characteristic analysis was performed using the SMAS in 300 IVF cycles using sperm from 270 men. At least three metaphase II oocytes were inseminated in C-IVF. The SMAS estimates were measured in semen and before insemination. This study was approved by the Ethics Committee of the Institutional Review Board (IRB) of Kinutani Women’s Clinic, Hiroshima, Japan. Participants/materials, setting, methods Motile sperm from at least 3 different fields were observed in each sample and their parameters were examined. First, we investigated the correlation between the normal fertilization rates and sperm motility characteristics. Next, the group comprising 300 cycles was classified into a group of a ≥ 60% normal fertilization (group 1: 229 cycles) and a group with &amp;lt;60% normal fertilization (group 2: 71 cycles) to assess the sperm characteristics described below. Main results and the role of chance The following indicators were evaluated: amplitude lateral head displacement (ALH), best cross frequency (BCF), curvilinear velocity (VCL), straight-line velocity (VSL), average path velocity (VAP), linearity (LIN=VSL/VCL), straightness (STR=VSL/VAP), and curvilinear path wobble (WOB=VAP/VCL).There were significant correlations between normal fertilization rates and semen parameters, including VCL (r = 0.014; P &amp;lt; 0.05) and ALH (r = 0.177; P &amp;lt; 0.05). However, there were no significant correlations for other parameters. Moreover, there were no significant correlations between normal fertilization rates and sperm parameters before insemination. The SMAS estimates in semen and sperm before insemination were compared between group 1 and group 2. There were no significant differences in most baseline characteristics across groups, except for the number of normally fertilized oocytes. However, there were significant differences in sperm WOB before insemination between group 1 and group 2 (0.589 vs. 0.572; P &amp;lt; 0.05). Sperm motility before insemination was more curvilinear in group 1 than in group 2. The optimal cut-off value for WOB of motile sperm before insemination for predicting≥60% normal fertilization was 0.587 (AUC was 0.60; P &amp;lt; 0.05). These results were suggested that the WOB of sperm before insemination is useful as an indicator in determining the selection of methods for fertilization in vitro. Limitations, reasons for caution The main limitations of this study are its retrospective design and small sample size. A prospective study is necessary to further clarify the prediction criteria. Wider implications of the findings This study suggests that the WOB of motile sperm before insemination may be used as one of the indicators to predict good fertilization rates after C-IVF. Trial registration number N.A.

P-215 Use of hyaluronan-based solution as an alternative to polyvinylpyrrolidone to improve blastulation in ICSI

Abstract Study question This study aimed to analyze whether hyaluronan as an alternative to polyvinylpyrrolidone (PVP) for sperm immobilization, aspiration, and injection into the ooplasm improves ICSI outcomes. Summary answer The use of hyaluronan solution as an alternative to PVP improves the blastocyst formation rate and good quality blastocyst formation rate compared to PVP. What is known already The PVP solution facilitates ICSI procedures, such as sperm immobilization, aspiration, and injection into the ooplasm. However, it has been reported that injection of a small amount of PVP along with the sperm into the ooplasm negatively affects subsequent embryo development. Hyaluronan is a natural component of the extracellular matrix of the cumulus-oocyte complex. Balaban et al. (2003) reported that hyaluronan can replace PVP during ICSI procedures in the early embryo transfer cycle without adversely affecting pregnancy outcomes. This study analyzed whether using hyaluronan-based solution for the ICSI procedure would improves blastocyst development compared with ICSI using PVP. Study design, size, duration This retrospective study analyzed clinical medical reports at Takahashi Women’s Clinic, Japan. We included 4002 oocytes retrieved from 411 patients under 39 years of age who underwent ICSI using autologous oocytes between December 2019 and August 2021. Of the oocytes used for ICSI, 1909 underwent sperm aspiration and injection into the ooplasm with hyaluronan (Sperm Slow; Origio), and 2093 oocytes with 7% PVP (NakaMedical). We did not perform preimplantation genetic testing-aneuploidy for any cycles. Participants/materials, setting, methods We used PVP droplets for sperm selection under 400× magnification in both groups. In hyaluronan-ICSI, the selected sperm were transferred to a hyaluronan droplet and washed three times. The sperm was then aspirated with hyaluronan, and ICSI was performed. In PVP-ICSI, all procedures were performed using PVP. The ICSI and embryo transfer outcomes were compared between hyaluronan-ICSI and PVP-ICSI by logistic regression analysis considering patient age, BMI, and basal level of anti-mullerian hormone. Main results and the role of chance Normal fertilization rates were 74.3% (1556/2093) in PVP-ICSI and 75.5% (1442/1909) in hyaluronan-ICSI. There was no significant difference in the normal fertilization rate between PVP-ICSI and hyaluronan-ICSI groups (p = 0.437, aOR:1.06, 95% CI: 0.92–1.22). We cultured 1323 2PN embryos in PVP-ICSI and 1237 2PN embryos in hyaluronan-ICSI until the blastocyst stage. Blastocyst formation rates were 48.1% for PVP-ICSI and 52.3% for hyaluronan-ICSI, and this difference was statistically significant (aOR, 1.20; 95% CI: 1.02–1.40; p = 0.024). Moreover, the good grade (Gardner criteria ≧BB) blastocyst formation rates were significantly higher in the hyaluronan-ICSI group (36.9% and 41.0%, aOR: 1.21, 95% CI: 1.03–1.42, p = 0.022). During the study period, we performed 163 and 169 cryo-thawed blastocyst transfer cycles in PVP-ICSI and hyaluronan-ICSI, respectively. The clinical pregnancy rate (50.9% vs. 54.4%, aOR: 1.19, 95% CI: 0.77–1.83, p = 0.443) and miscarriage rate (19.3% vs. 13.0%, aOR: 0.66, 95% CI: 0.3–1.44, p = 0.295) after embryo transfer were not significantly different between PVP-ICSI and hyaluronan-ICSI. Limitations, reasons for caution The study was conducted at a single IVF center, and the oocytes included in this study were collected from patients aged &amp;lt; 39 years. Embryo transfer result is based on ongoing pregnancy, while the live birth data for all pregnancies are not yet available. Wider implications of the findings Hyaluronan facilitates ICSI procedures such as sperm immobilization, aspiration, and injection. Moreover, the hyaluronan improves blastocyst development. The present study indicates that using hyaluronan as an alternative to PVP during the ICSI procedure is recommended. Trial registration number not applicable