Embryo Euploidy Rates Research Articles

P-547 Impact of SARS-COV-2 infection on embryo euploidy rate: a retrospective cohort study of 498 PGT-A cycles in China

Abstract Study question Does SARS-CoV-2 infection before oocyte retrieval have any impacts on embryo euploidy rate? Summary answer SARS-CoV-2 infection before oocyte retrieval had no detrimental impact on embryo euploidy rate, indicating that cycle cancellation due to SARS-CoV-2 infection may not be necessary. What is known already SARS-CoV-2 infection has significant effects on the health of people worldwide and poses a serious threat to reproductive health. According to the previous studies, SARS-CoV-2 infection had negative impacts on the blastocyst formation rate, top-quality embryos rate. However, the association between the infection and the embryo euploidy rate is indeterminate. It is still worth exploring the impact of SARS-CoV-2 infection on embryo euploidy rate. Especially this is the key question to be addressed for providing appropriate counseling to couples seeking for preimplantation genetic testing for aneuploidy (PGT-A) treatments during the ongoing pandemic. Study design, size, duration This was a retrospective cohort study of 498 couples who underwent PGT-A cycles with next-generation sequencing technology from November 1st, 2022 to June 30th, 2023 at a tertiary-care medical center in China. Participants/materials, setting, methods Women tested positive for SARS-CoV-2 before oocyte retrieval were included in the COVID-19 group, while patients who were asymptomatic and tested negative during the same period were included in the non-COVID-19 group. The primary outcome was the euploidy rate. Subgroup analysis was conducted according to the time interval between infection and oocyte retrieval. Univariate and multivariate logistic regression models were applied to adjust for potential confounders. Main results and the role of chance A total of 278 and 220 women were included in the COVID-19 and non-COVID-19 group, respectively. In the COVID-19 group, the maturated oocyte rate (79.5% vs. 83.7%, p＜0.001), number of total viable embryos (2 vs. 3, p=0.038), and number of euploid blastocyst (1 vs. 1, p=0.028) were significantly decreased compared with non-COVID-19 group. The euploidy rate (38.5% vs.40.3%, p = 0.451), aneuploidy rate (45.0% vs. 43.8%, p=0.616), and mosaicism rate (16.5% vs. 15.9%, p=0.746) were all comparable between the two groups. Results of multivariate logistic regressions revealed no association between SARS-CoV-2 infection and euploidy rate, aneuploidy rate, or mosaicism rate. Similarly, no association was detected between the time interval of infection and oocyte retrieval (≤30 days, 31-60days, 61-90 days, 91-120 days, or ≥ 120 days prior to oocyte retrieval) and euploidy rate. Limitations, reasons for caution The retrospective nature of the study and limited sample size for subgroup analysis limits the generalizability of the findings. Other potential confounders, such as the level of SARS-CoV-2 antibody, symptoms of infection, are needed to be taken into consideration as well. Large-scale prospective studies are warranted to confirm the conclusion. Wider implications of the findings Our study demonstrated that SARS-CoV-2 infection before oocyte retrieval had no detrimental impact on embryo euploidy rate, indicating that cycle cancellation due to SARS-CoV-2 infection may not be necessary. Trial registration number Not Applicable

P-074 The effect of sperm from male patients who use electronic cigarettes on the euploidy ratio of embryos during in vitro fertilization procedures

Abstract Study question What is the effect of electronic cigarettes as a substitute for regular cigarettes on sperm and subsequent embryo development in IVF patients? Summary answer E-cigarettes are just as harmful to male fertility as regular cigarettes, reduce the euploidy rate of the embryo, and have negative effects on pregnancy. What is known already Many harmful substances from smoking are known to negatively affect sperm development and function. It causes high elevations in sperm DNA fragmentation (SDF), which are associated with embryo euploidy rates. Recently, the use of e-cigarettes is increasing as an alternative to regular cigarettes because they are less harmful and safer. However, several animal studies have shown that e-cigarettes are just as harmful as regular cigarettes and can have negative effects on sperm and fertility. Preliminary data showed that SDF increased with e-cigarettes compared to regular cigarettes. Therefore, we would like to find out whether e-cigarettes affect embryo euploidy. Study design, size, duration In this study, from January 2022 to December 2023, 1,725 cycles of PGT-A (PGT for aneuploidy) were analyzed among the patients of IVF. The average ages of men and women were 40.0±4.6 and 38.7±4.0. A total of 4,796 embryos were analyzed (2.8 ± 2.0 per case) for euploidy rates. Participants/materials, setting, methods Blastocyst biopsy and PGT-A were performed, and embryo euploidy rates was confirmed. In male, the number of no smoking IVF cycles was 1061 and the number of embryo biopsies was 2.9 ± 2.1. The number of IVF cycles using regular cigarettes is 370, and the number of embryo biopsies is 2.8 ± 2.1. The number of IVF cycles using e-cigarettes was 294, and the number of embryo biopsies was 2.5 ± 1.6. Main results and the role of chance Based on the total female age, the embryo euploidy rate was nonsmoking (27.3%), regular cigarettes (23.6%), and e-cigarettes (18.8%). Among women under 38 years of age, e-cigarettes (29.9%) had a lower euploidy rate than regular cigarettes (37.4%), and nonsmoking (38.2%) had the highest rate. Among women over 38, the euploidy rate dropped significantly compared to those under 38, including nonsmoking (18.9%), but it was also lower for e-cigarettes (11.6%) than regular cigarettes (17.7%). Based on female age under 38 years old, the embryo euploidy rate among male age under 39 years old was nonsmoking (39.0%), regular cigarettes (38.3%), and e-cigarettes (31.7%). And among men over 40, the group with the lowest embryo euploidy rate was those who used e-cigarettes (no smoking (35.0%), regular cigarettes (32.6%), e-cigarettes (27.7%)). Based on the age of women over 38 years old, the euploidy rate among men under 39 years of age is nonsmoking (23.9%), regular cigarettes (21.6%), and e-cigarettes (12.7%). And the euploidy rate among men over 40 years of age is nonsmoking (16.8%), regular cigarettes (16.5%), and e-cigarettes (11.8%). nonsmoking is essential to maintain a high euploidy rate in embryos, and e-cigarettes may have a more adverse effect on pregnancy than regular cigarettes. Limitations, reasons for caution Because there are various types of e-cigarettes, it is expected that the degree of harmfulness will vary, but the number of patients was not sufficient to conduct research on each type of e-cigarette. Therefore, it is believed that the above problems should be solved through additional research. Wider implications of the findings This study is a study that links the harmful effects of cigarettes and the polyploidy of embryos. Electronic cigarettes, used as an alternative to regular cigarettes, may reduce embryo euploid production and cause recurrent pregnancy failure. Therefore, it is believed that electronic cigarettes should also be managed appropriately. Trial registration number not applicable

P-058 The role of sperm selection in improving embryo euploidy rate: a comparison between Swim-up and Microfluidic Sperm Selection

Abstract Study question Does Microfluidic Sperm Selection (MSS) improve the blastocysts’ development rate and the euploidy rate compared to Swim-up sperm selection? Summary answer MSS doesn’t improve the rate of blastocyst development and euploidy compared to Swim-up. Furthermore, in women age ≥ 37 years D6 aneuploid blastocyst rate increases What is known already Sperm defects have been shown to have a negative impact on embryo quality and development. MSS was introduced as an alternative to traditional centrifugation-based sperm preparation techniques to efficiently isolate highly motile and healthy spermatozoa. Centrifugation has been suggested to compromise membrane integrity, motility, mitochondrial energy production, and DNA structure. Less DNA fragmentation and ROS formation was observed in spermatozoa isolated with MSS compared to standard techniques. Our study aims to analyze whether MSS also improves biological outcomes. Study design, size, duration In a prospective study of 51 ICSI/PGT-A cycles performed between September 2022 and November 2023, freshly ejaculated semen sample was divided into two aliquots and processed by Swim-up and MSS. Split sperm sample was used to inject siblings’ mature oocytes (249 vs 274 oocytes in Swim-up vs MSS groups). Inclusion criteria: retrieved MII oocytes ≥8, oligoteratozoospermic and normozoospermic semen. Results were analyzed according to women’s age <37 years (21 cycles) and ≥37 years (30 cycles). Participants/materials, setting, methods Spermatozoa were selected using either conventional Swim-up or a microfluidics device (ZyMōtTM). Post treatment sperm parameters (sperm count; total sperm motility; progressive motility and hyperactivated motility) were examined for both. Embryo culture was performed in Irvine Continuous Culture-NX media until day5 or day6 and biopsy was performed at the blastocyst stage by next generation sequencing. Primary endpoint was fertilization rate, blastocyst development and usable blastocysts rate. Chi-square analysis was performed and p < 0.05 was considered significant Main results and the role of chance There is no significant differences overall between Swim-up and MSS in fertilization rate (73% vs 75%), blastocyst development rate (47% vs 54%) and ploidy rate: aneuploid (47% vs 50%), usable blastocyst (i.e. euploid+mosaics) (53% vs 50%). When considering female age, no significant difference is observed in the primary endpoints in the <37 years group. However, in women aged ≥37 years, the rate of blastocyst development is similar in the two sperm preparation groups, but a significant difference is observed in the timing of blastocyst development: The percentage of blastocysts formed on day 6 in the MSS group is significantly higher than in the Swim-up group (43% vs. 19% p < 0.05). In addition, the aneuploidy rate of blastocysts on day 6 is also significantly higher than in the Swim-up group (79% vs. 50% respectively, p < 0.05). The concentration of sperm recovered post-treatment was similar in Swim-up vs MSS (23 x106 vs 19 x106); total motility and progressive motility were similar in Swim-up compared to MSS (92.7% vs. 95.3% and 85% vs. 87% respectively). Only hyperactivated motility assessed with the SCASCOPE CASA System (Microptic) was significantly higher in MSS vs Swim-up 41% vs 4% respectively (p < 0.0005). Limitations, reasons for caution Our study included couples with different infertility diagnosis and semen parameters. Further studies with an increased number of patients may provide more comprehensive data to better evaluate biological and clinical outcomes to eventually suggest the use of MSS in laboratory routines Wider implications of the findings Recently, some studies reported that microfluidic-sorted sperm showed significantly less ROS formation and DNA fragmentation compared to those treated with conventional Swim-up method. Nevertheless, our data does not currently show the expected increase in embryo euploidy rate to justify the routine use of these expensive devices in male infertility treatment Trial registration number non applicabile

P-612 Euploidy rates between cycles triggered with gonadotrophin-releasing hormone agonist (GnRHa), human chorionic gonadotrophin (hCG) and dual trigger

Abstract Study question Is there any difference in euploidy rates between in vitro fertilization (IVF) cycles triggered with GnRHa, hCG or dual trigger? Summary answer There is no difference in the euploidy rate in preimplantation genetic testing for aneuploidy (PGT-A) cycles according to the trigger method used. What is known already The use of PGT-A has shown that the success of meiosis in the IVF laboratory is inversely correlated with maternal age. However, other uninvestigated factors involved in the IVF process may interfere with normal oocyte physiology. Since the final maturation obtained after ovarian stimulation depends on the use of drugs to promote the resumption of meiosis, it is worth investigating whether the triggering method could impact on oocyte and embryo euploidy. This study aims is to evaluate differences in embryo euploidy rates between PGT-A cycles triggered with GnRHa, hCG or dual trigger (GnRHa + hCG). Study design, size, duration This is a single-center retrospective study of 934 patients who underwent 1085 PGT-A cycles by Next-Generation Sequencing (NGS) from January 2017 to December 2023, using one of three possible trigger methods to induce final oocyte maturation: GnRHa, hCG, or dual trigger. The couples included in the study had normal karyotypes and no severe male factor. Participants/materials, setting, methods Data on the number of oocytes retrieved, ratio of metaphase II (MII) oocytes and fertilization rate, blastocyst development and embryo euploidy were assessed and compared between the GnRHa, hCG and dual trigger treatments. A secondary analysis was performed to evaluate data stratified by patient age. Main results and the role of chance A total of 1085 PGT-A cycles involving 934 patients (38.07 ± 3.01 years) and 2803 biopsied blastocysts were included in the analysis. Dual trigger was used in 726 (77.73%) cycles, GnRHa in 287 (26,4%) cycles and hCG in 72 (7.71%) cycles. The number of oocytes retrieved was significantly higher in the GnRHa group (GnRHa: 18.72 ± 8.59; hCG: 9.88 ± 5.64; dual trigger: 10.58 ± 5.65; p < 0.001) with similar maturation rate (GnRHa: 77.46%; hCG: 74.58%; dual trigger: 76.03%; p = 0.789), fertilization rate (GnRHa: 78.12%; hCG: 76.37%; dual trigger: 79.12%; p = 0.168), blastulation rate (GnRHa: 61.03%; hCG: 58.32%; dual trigger: 58.73%; p = 0,629), high-quality blastocyst rate (GnRHa: 27,88%; hCG: 33,67%; dual trigger: 27,88%; p = 0.543) and embryo euploidy per biopsied embryo (GnRHa: 42.48%; hCG:45.49%; dual trigger:46.98%; p = 0.112). Analysis of the data stratified by age showed similar rates of euploidy rates with the three trigger methods in women younger than 41 years (<35 years: GnRHa 66.50%; hCG 66.67%; dual trigger 56.97% p = 0.290; 35-37 years: GnRHa 49.45% hCG 51.04%; dual trigger 50.23%; p = 0.988; 38-40 years: GnRHa 40.93%; hCG 41.27%; dual trigger 39.13% p = 0.257). However, in patients ≥41 years old the euploidy rate was statistically higher in the GnRHa group (GnRH 31.51%; hCG 18.75%; dual trigger 16.89%; p = 0.032). Limitations, reasons for caution The main limitations of the study are: its retrospective design, the possibility that the trigger method was chosen based on the ovarian response and the limited number of patients in both the hCG trigger and women ≥ 41 years groups. Wider implications of the findings Our results suggest that triggering with GnRHa, hCG or dual trigger leads to comparable euploidy rates. Triggering with GnRHa should be recommended in older patients since it has been associated with a higher rate of embryonic euploidy. Trial registration number Not Applicable

P-050 Microfluidic chip-sorted spermatozoa selection yields embryo euploidy rate and reproductive outcomes similar to density-gradient centrifugation

Abstract Study question Does microfluidic chip-sorted spermatozoa selection (MSS) improve good quality blastocyst (GQB) and embryo euploidy rates compared to density gradient centrifugation (DGC)? Summary answer The use of MSS before intracytoplasmatic sperm injection (ICSI) does not improve good quality blastocist and euploidy rates comparing to DGC. What is known already The integrity of sperm DNA plays a crucial role in embryonic development. Conventional sperm selection methods, such as DGC, may potentially damage sperm DNA due to oxidative stress generated during centrifugation. One of the recent advances in sperm selection techniques is the microfluidic chamber, which, requiring less processing time and eliminating the need for centrifugation, allows for the selection of sperm with better mobility, morphology, and reduced DNA damage. Consequently, improved GQB and euploid embryo rates, as well as enhanced reproductive outcomes, are expected. Study design, size, duration Retrospective study of 1739 blastocysts from 404 couples undergoing 544 autologous cycles of ICSI/PGT-A (preimplantation genetic testing for aneuploidies) from January 2020 to December 2022. Those patients with sperm concentration <5 × 106/mL or motility <10%, altered karyotype, women aged > 43 years, uterine anomalies and thrombophilia were excluded. Besides comparing euploidy rates and blastocyst formation, reproductive outcomes were analyzed and compared. Participants/materials, setting, methods According to the sperm selection technique, two groups were established. One group underwent MSS (ZyMōt ICSI® or ZyMōt Multi®), resulting in 295 blastocysts from 96 cycles. It was compared with a control group (DGC) including 1444 blastocysts from 448 ICSI/PGT-A cycles. PGT-A was performed in day 5/6 using Next Generation Sequencing (NGS) technology. For a 95% confidence interval, statistical significance was considered at p < 0.05. Chi-square analysis was used for the statistical evaluation. Main results and the role of chance Both groups were comparable in terms of demographic data, including age, sperm analysis parameters, and cycle characteristics. There were no significant differences between the DGC and MSS groups in the women’s age (mean ± SD) (37.9 ± 3.9 vs. 38 ± 3.9 years, p = 0.99) or men’s age (38.4 ± 7.1 vs. 39.8 ± 5.9 years, p = 0.11). No significant differences were found between the DGC and MSS groups in terms of GQB rate (49.3% vs. 50.7%, p = 0.55), euploidy rate (35.8% vs. 32.2%, p = 0.23), fertilization rate (69.5% vs. 69.5%, p = 0.99), clinical pregnancy rate per transfer (63% vs. 76%, p = 0.13), clinical miscarriage (9% vs. 21%, p = 0.33), and live birth rate per transfer (44% vs. 37%, p = 0.42). The results also showed a higher number of euploid embryos per initiated cycle in the DGC group (1.154) compared to the MSS group (0.989). Despite this difference, the results were not statistically significant (p > 0.05). Limitations, reasons for caution The limitations of this study include the potential impact of selection bias, as it is a retrospective study in which groups were not randomly assigned. Additionally, the low number of blastocysts in the MSS group and the potential bias of DNA fragmentation, which was not evaluated in the study. Wider implications of the findings Both methods demonstrated similar effectiveness. While MSS could potentially be more effective in samples with high DNA fragmentation and they feature a user-friendly application, its major limitation lies in its challenging application with samples having a low concentration of motile sperm. Specific semen samples may need different sperm selection methods. Trial registration number Not Applicable

P-009 Impact of sperm preparation methods on embryo development rate and embryo ploidy status: ICSI egg donation as a model

Abstract Study question Do different sperm separation techniques result in distinguishable contributions to embryo development rates and ploidy status? Summary answer CA0 sperm separation device, devoid of centrifugation, yielded a higher euploidy rate than conventional swim-up (50% vs. 74%, p = 0.02), notably observed in abnormozoospermic cases. What is known already Sperm DNA fragmentation (SDF) emerges as a key paternal risk factor associated with embryo aneuploidy, increasing the risk of ART failures. Factors like advanced male age, prolonged ejaculatory abstinence, and the detrimental effects of centrifugation during sperm separation jeopardize sperm DNA integrity. A recent centrifugation-free sperm separation device (CA0), employing live sperm sorting technology, exhibits efficiency in producing highly motile sperm with minimized SDF. This retrospective cohort study solely focusing on ICSI egg donation cycles to assess the influence of sperm separation methods (conventional swim-up vs. CA0) on early embryo development, and embryo ploidy. Study design, size, duration This retrospective study included 83 couples who underwent ICSI egg donation cycles at Chachava Clinic, Tbilisi, Georgia. Thirty-three semen specimens were processed using conventional swim-up (SU) and 427 Mii were inseminated. Forty-four semen samples were separated by CA0 and 519 Mii were inseminated. Clinical outcome measures included fertilization rate (number of 2PN over number of inseminated oocyte), good embryo rate (embryo number with more than 8 cells over inseminated oocyte), and euploidy rate using PGT-A. Participants/materials, setting, methods Male age and basic semen parameters via CASA (CEROS II, Hamilton-Thorne) were recorded. SU involved layering 1 mL neat semen to 1.2 mL HTF medium, inclining the tube at 45° for 60 min at 37 °C, collecting 1 mL uppermost, adding 2 mL HTF for centrifugation at 500×g for 5 min, and resuspending sperm pellet with 0.5 mL HTF. Semen samples were sorted using LensHooke® CA0 device following the manufacturer’s instructions. Main results and the role of chance In comparing demographic characteristics between SU and CA0 arms, CA0 exhibited higher median total motility (SU: 40% vs. CA0: 54%, p = 0.02), with no significant differences in female age and Mii oocyte count. This study assessed the impact of sperm separation methods on early embryo development, finding no significant differences in fertilization rate (SU: 79% vs. CA0: 77%) and good embryo rate (SU: 64% vs. CA0: 62%). However, CA0 demonstrated a significantly higher euploidy rate than SU (SU: 59% vs. CA0: 69%, p = 0.03). Further analysis within subpopulations, including normozoospermic and asthenozoospermic (total motility <40%), revealed equivalent euploidy rates between SU and CA0 for normospermic specimens (SU: 66% vs. CA0: 68%). However, a significant differentiation in euploidy rate emerged for asthenozoospermic cases (SU: 59% vs. CA0: 74%, p = 0.02). Specifically, comparing outcomes in normozoospermic, a significant reduction was observed in asthenozoospermic cases using SU (66% vs. 50%, p = 0.02), whereas CA0 provided improved euploidy rates regardless of semen quality (68% for normozoospermic and 74% for asthenozoospermic). In conclusion, this study suggests that the non-invasive live sperm sorting device enhances embryo euploidy rates, serving as a reliable sperm preparation method, ensuring selection consistency despite sample variations. Limitations, reasons for caution This study’s limitations include its retrospective nature and single-center trial design. To more effectively assess CA0 efficiency, a randomized controlled trial targeting diverse patient eligibility criteria is warranted. Wider implications of the findings Attributed to the improved embryo euploidy with CA0, we anticipate enhanced ART outcomes, such as increased embryo transfer rate and prevention of ART failures related to embryo aneuploidy. Additionally, CA0 offers practical benefits by streamlining and standardizing the sperm separation process with only three steps involved. Trial registration number not applicable

The comparision among euploidy of preimplantation blastocysts in different controlled ovary stimulation (COH) protocols.

To compare differences in euploidy rates for blastocysts in preimplantation genetic testing for aneuploidy (PGT-A) cycles after gonadotropin-releasing hormone agonist (GnRH-a) long and short protocols, GnRH-antagonist (GnRH-ant) protocol, progestin-primed ovarian stimulation and mild stimulation protocols, and other ovary stimulation protocols. This was a retrospective cohort study from the Assisted Reproductive Medicine Department of Shanghai First Maternity and Infant Hospital. A total of 1657 PGT-A cycles with intracytoplasmic sperm injection after different controlled ovary hyperstimulation protocols were analyzed, and a total of 3154 embryos were biopsied. Differences in euploidy rate per embryo biopsied, embryo euploidy rate per oocyte retrieved and cycle cancellation rate were compared. For the PGT-A cycles, the euploidy rate per embryo biopsied was lower in the GnRH-ant protocol than in the GnRH-a long protocol (53.26 vs. 58.68%, respectively). Multiple linear regression showed that the GnRH-ant protocol was associated with a lower euploidy rate per embryo biopsied (β = -0.079, p = 0.011). The euploidy rate per embryo biopsied was not affected by total gonadotropin dosage, duration of stimulation and number of oocytes retrieved. The embryo euploidy rate per oocyte retrieved was similar in all protocols and was negatively correlated with the total number of oocytes retrieved (β = -0.003, p = 0.003). Compared with the GnRH-a long protocol, the GnRH-ant protocol was associated with a lower euploidy rate per embryo biopsied. The total gonadotropin dosage, duration of stimulation and number of oocytes retrieved did not appear to significantly influence euploidy rates.

The impact of microfluidics sperm processing on blastocyst euploidy rates compared with density gradient centrifugation: a sibling oocyte double-blinded prospective randomized clinical trial

ObjectiveTo compare euploidy rates among blastocysts created from sibling oocytes injected with sperm processed by microfluidics or by density gradient centrifugation. DesignSibling oocyte randomized controlled trial. SettingSingle university-affiliated infertility practice. Patient(s)106 patients aged 18-42 undergoing fresh in vitro fertilization (IVF) cycles with preimplantation genetic testing (PGT) between January 2021 to April 2022 contributed 1442 mature oocytes which were injected with sperm processed via microfluidics or density gradient centrifugation. Intervention(s)Sperm sample divided and processed via a microfluidics device and density gradient centrifugation for injection into sibling oocytes. Main outcome measure(s)Primary outcome was the embryo euploidy rate. Secondary outcomes included the fertilization, high quality blastulation, and ongoing pregnancy rates. Result(s)The blastocyst euploidy rate per mature oocyte was not significantly different in the study group compared to the control group (22.9% versus 20.5%, p = 0.60). The blastocyst euploidy rate per biopsied embryo was also similar between the two groups (53.0% versus 45.7%, p = 0.34). However, the fertilization rate per mature oocyte injected was found to be significantly higher in the study group compared to the control group (76.0% versus 69.9%, p = 0.03). The high quality blastulation rate per mature oocyte injected was similar between the two groups, as was the total number of embryos frozen. There were no differences in the number of participants with no blastocysts for biopsy or the number of participants with no euploid embryos between the two groups. Among the male factor infertility and recurrent pregnancy loss subgroups, there were no differences in euploidy rates, fertilization rates, blastulation rates, or total numbers of blastocysts frozen, although the study was underpowered to detect these differences. Seventy-seven patients underwent frozen embryo transfer; there were no significant differences in pregnancy outcomes between the two groups. Conclusion(s)Microfluidics processing did not improve embryo euploidy rates compared to density gradient centrifugation in this sibling oocyte study, although fertilization rates were significantly higher.

Optimal antimüllerian hormone levels in oocyte donors: a national database analysis

To study the relationship between high antimüllerian hormone (AMH) levels in oocyte donors and embryo development and pregnancy outcomes among donor oocyte recipients. Retrospective cohort study. Donor Egg Bank Database. Patients undergoing invitro fertilization using vitrified donor oocytes from 35 invitro fertilization centers in the United States between 2013 and 2021. For each recipient, the first oocyte lot that was received with a planned insemination and embryo transfer (ET) was included. Oocyte donor-recipient cycles. Ongoing pregnancy rate (OPR) per ET. A total of 3,871 donor oocyte-recipient thaw cycles were analyzed. On the basis of donor AMH serum concentration, cycles were stratified into the high AMH group (AMH ≥5 ng/mL; n = 1,821) and the referent group (AMH <5 ng/mL; n = 2,050). Generalized estimating equation models were used to account for donors that contributed more than one lot of oocytes. The number of usable embryos per lot (median [interquartile range]) was significantly increased in the high AMH group (2 [2-4]) compared with the referent group (2 [1-3]) (relative risk [RR] 1.06; confidence interval [CI] 1.01-1.12). Among recipients with a planned ET, there was no difference in OPR between the high AMH group (45.4%) and the referent group (43.5%) (RR 1.04; 95% CI 0.94-1.15). Among preimplantation genetic testing for aneuploidy cycles, the embryo euploidy rate per biopsy was similar at 66.7% (50%-100%) in both groups (RR 1.04; CI 0.92-1.17). The OPR per euploid ET among patients who used preimplantation genetic testing for aneuploidy was also comparable, at 52% in the high AMH group and 54.1% in the referent group (RR 0.95; CI 0.74-1.23). This large national database study observed that there was no association between a high level of AMH (≥5 ng/mL) in oocyte donors and an OPR in the recipient after the first ET. On the basis of these findings, recipients and physicians can be reassured that oocyte donors with a high AMH level can be expected to produce outcomes that are at least as good as donors with an AMH level (<5 ng/mL).

The impact of the number of previous miscarriages on embryo euploid rate and pregnancy outcomes in patients with unexplained recurrent pregnancy loss

Objective: To investigate the impact of the number of previous miscarriages on embryo euploid rate and pregnancy outcomes after preimplantation genetic testing for aneuploidies (PGT-A) in patients with unexplained recurrent pregnancy loss (uRPL). Methods: A retrospective cohort study was conducted. 799 women with uRPL who underwent PGT-A for the first time between January 2015 and December 2021 at the Reproductive center of Shandong University were enrolled. These patients were divided into three groups according to the number of previous miscarriages (2, 3, and≥4). Stratified analysis was conducted according to female age (≤37 years and>37 years). The embryo euploidy rate, good-quality blastocyst formation rate, cumulative live birth rate, and cumulative clinical pregnancy loss rate of three groups were compared in younger and older patients, respectively. Meanwhile, the cumulative live birth rate, clinical pregnancy loss rate, and embryo euploidy rate were analyzed by multivariate logistic regression analysis. Results: Patients' age was (34.7±5.1) years old. In the three groups with 2, 3 and ≥4 previous miscarriages, there was no significant difference in the embryo euploidy rate between groups in the younger [48.9% (539/1 103), 50.6% (354/700) and 52.1% (152/292), P=0.567] and older [26.2% (103/393), 28.8% (55/191) and 20.5% (16/78), P=0.377] age population. Compared with 2 and 3 previous miscarriages, the cumulative live birth rate was significantly decreased [52.6% (153/291), 52.8% (93/176) and 34.3% (25/73), P=0.014] and the cumulative clinical pregnancy loss rate was significantly increased [15.8% (31/196), 15.3% (18/118) and 46.9% (23/49), P<0.001] in younger women with ≥4 miscarriages. After adjusting for maternal age, BMI, AMH, endometrial thickness on hCG trigger day and antral follicle count, the number of previous miscarriages ≥4 was a relevant factor for cumulative live birth rate (OR=0.461, 95%CI: 0.263-0.807, P=0.007) and the cumulative clinical pregnancy loss rate (OR=4.382, 95%CI: 2.165-8.873, P<0.001) in younger patients, but it was not significantly correlated with the cumulative live birth rate, cumulative clinical pregnancy loss rate and embryo euploidy rate in patients with advanced age. Conclusion: In uRPL patients,≥4 previous miscarriages decreased cumulative live birth rate and increased cumulative clinical pregnancy loss rate in women aged≤37 years old.

Compared with conventional insemination, intracytoplasmic sperm injection provides no benefit in cases of nonmale factor infertility as evidenced by comparable euploidy rate

To evaluate if differences in euploidy rates exist between intracytoplasmic sperm injection (ICSI) and conventional insemination (CI) in non-male factor infertility cases. Retrospective cohort study. Single, academically affiliated infertility center in the U.S. A total of 3,554 patients who underwent in-vitro fertilization (IVF) cycles from January 2014 to December 2021. All cycles that had pre-implantation testing for aneuploidy (PGT-A) performed by trophectoderm biopsy and had a post preparation sperm concentration >4 million total motile sperm per milliliter were included. The primary outcome was embryo euploidy rate per embryo biopsied in the ICSI versus CI group. Secondary outcomes included fertilization rate and number of embryos biopsied. Generalized estimating equations with a Poisson distribution was used to estimate the euploid rate ratio (with total embryos biopsied as an offset), while accounting for multiple retrievals per patient. To adjust for confounding, a propensity score model was fit for ICSI using 14 baseline female and male characteristics. Oocytes retrieved and number of embryos biopsied were similar in both groups, while fertilization rate per oocyte retrieved was significantly lower with ICSI (0.64 vs 0.66, p=0.04). The proportion of euploid embryos in the ICSI group was significantly lower when compared to CI (0.47 vs 0.52), with a euploid rate ratio of 0.89 (p<0.001). Interestingly, when accounting for variation in PGT reference laboratories over the study time period, adjusting for date of procedure did not change the relationship between ICSI and euploid rate (RR=0.89, p<0.001); however, after adjusting for PGT reference laboratory, the relationship between ICSI and euploid rate was no longer significant (RR=0.97, p=0.07). In the setting of non-male factor infertility, ICSI resulted in a lower fertilization rate and an 11% lower embryo euploid rate compared to CI. Although the data is not statistically significant when adjusted for PGT reference laboratory, we can still conclude that ICSI does not provide any benefit. This data supports the recommendation that CI should be the preferred methodology of fertilization in non-male factor infertility cases.

Can a sperm selection technique improve embryo ploidy?

Spermatozoa with the highest motility retain a superior genomic integrity, and elevated sperm chromatin fragmentation (SCF) has been linked to a lower ability of the conceptus to develop and implant. Therefore, the utilization of a sperm selection method, such as microfluidic sperm selection (MFSS), is capable of reducing the SCF by yielding the most motile fraction of spermatozoa with the highest embryo developmental competence. What remains unclear, however, is the causal mechanism that linksSCF to an impaired embryo development. To identify a relationship between SCF and an unexpectedly high proportion of embryo aneuploidy, while addressing treatment options. We identified couples with a high incidence of embryo aneuploidy in a previous intracytoplasmic sperm injection (ICSI) cycle with pre-implantation genetic testing for aneuploidy (PGT-A), utilizing spermatozoa selected by density gradient (DG). Terminal deoxynucleotidyl dUTP transferase nick-end labeling (TUNEL) and neutral Comet assays were carried out on the semen specimens to assess total SCF and double-stranded DNA (dsDNA) fragmentation, respectively. These couples underwent subsequent ICSI/PGT-A cycles with MFSS. Total SCF and dsDNA fragmentation were compared between the two sperm selection methods. Embryo aneuploidy, implantation, clinical pregnancy, delivery,and pregnancy loss rates were compared between the couples' historical DG and subsequent MFSS cycles. In 57 couples undergoing 71 ICSI/PGT-A cycles, where DG sperm selection was carried out, a high incidence of aneuploid embryos (74.7%) resulted in poor implantation and no viable pregnancies. Testing for SCF, inclusive of dsDNA breaks, evidenced a SCF of 26.2% and dsDNA break of 3.6% in the raw specimen, that decreased to 18.0% (p < 0.001) and 3.1%, respectively, in the DG processed specimen. Following MFSS, total SCF and dsDNA fragmentation decreased to 1.9% and 0.3%, respectively (p<0.001). The embryo euploidy rate remarkable improved from 25.3% in the DG cycles to 42.9% in the MFSS cycles (p<0.001). The 6.7% implantation rate in the DG cycles increased to 65.5% in the MFSS cycles (p<0.001). Similarly, the clinical pregnancy rate rose from 10.5% (DG) to 64.6% (MFSS), resulting in a 62.5% delivery rate (p<0.001). In couples with a relatively young female partner with a negative infertility workup, and a male partner with semen parameters adequate for ICSI, presenting with a high rate of embryo aneuploidy, an additional subtle male factor component may be the culprit. Thus, it is crucial to assess the SCF and test for the dsDNA breaks, which can eventually contribute to embryo chromosomal abnormalities. Given the inverse relationship between SCF and motility, a selection of the most motile gamete by MFSS enhanced the proportion of spermatozoa with an intact genome, contributing to the generation of moreeuploid embryos that are capable of implanting and yielding increased term pregnancies.

Comparison of blastocyst euploidy rates following luteal versus follicular phase stimulation in a GnRH antagonist protocol: a prospective study with repeated ovarian stimulation cycles.

Is there any difference in the mean number of euploid embryos following luteal phase start (LS) and follicular phase start (FS) of ovarian stimulation? The mean number of euploid blastocysts is equivalent independent of whether the inseminated oocytes are derived from FS or LS. Starting ovarian stimulation at any time of the cycle ('random-start') is commonly used for emergency fertility preservation in cancer patients. A few retrospective studies have been published evaluating LS in women undergoing ovarian stimulation in the context of IVF, but there is a lack of robust data on the comparative efficacy of LS versus FS.Although 'random start' is commonly used in cancer survivors, few retrospective and uncontrolled studies have been published evaluating luteal phase stimulation in women undergoing ovarian stimulation in the context of IVF. Owing to this evident lack of robust data on the efficacy of LS, guidelines typically recommend the LS approach only for medical reasons and not in the context of IVF. This is a prospective, equivalence study, with repeated stimulation cycles, conducted between May 2018 and December 2021. Overall, 44 oocyte donors underwent two identical consecutive ovarian stimulation cycles, one initiated in the FS and the other in the LS. The primary outcome of the study was to evaluate whether FS and LS in the same patient would result in equivalent numbers of euploid embryos following fertilization of oocytes with the same sperm sample. Overall, 44 oocyte donors underwent two consecutive ovarian stimulation protocols with 150 μg corifollitropin alpha followed by 200 IU recombinant FSH (rFSH) in a fixed GnRH antagonist protocol. The only difference between the two cycles was the day of initiation of ovarian stimulation, which was in the early follicular phase (FS) in one cycle, and in the luteal phase (LS) in the other. Forty-four oocyte recipients participated in the study receiving a mean of six metaphase II (MII) oocytes from each stimulation cycle (FS and LS). All MIIs were inseminated with the corresponding recipient's partner sperm (which had been previously frozen) or donor sperm, in order to safeguard the use of the same sample for either the FS or LS. Following fertilization and blastocyst culture, all generated embryos underwent genetic analysis for aneuploidy screening (preimplantation genetic testing for aneuploidy). FS resulted in a significantly shorter duration of ovarian stimulation (difference between means (DBM) -1.05 (95% CI -1.89; -0.20)) and a lower total additional dose of daily rFSH was needed (DBM -196.02 (95% CI -319.92; -72.12)) compared with LS. The donors' hormonal profile on the day of trigger was comparable between the two stimulation cycles, as well as the mean number of oocytes (23.70 ± 10.79 versus 23.70 ± 8.81) (DBM 0.00 (95% CI -3.03; 3.03)) and MII oocytes (20.27 ± 9.60 versus 20.73 ± 8.65) (DBM -0.45 (95% CI -2.82; 1.91)) between FS and LS cycles, respectively. Following fertilization, the overall blastocyst formation rate was 60.70% with a euploid rate of 57.1%. Comparisons between the two stimulation cycles did not reveal any significance differences in terms of fertilization rates (71.9% versus 71.4%), blastocyst formation rates (59.4% versus 62%) and embryo euploidy rates (56.9 versus 57.3%) for the comparison of FS versus LS, respectively. The mean number of euploid blastocysts was equivalent between the FS (1.59 ± 1.30) and the LS (1.61 ± 1.17), (DBM -0.02 (90%CI -0.48; 0.44)). The study was performed in young, potentially fertile oocyte donors who are patients with high blastocyst euploidy rates. Although results may be extrapolated to young infertile women with good ovarian reserve, caution is needed prior to generalizing the results to infertile women of older age. The current study provides evidence that initiation of ovarian stimulation in the luteal phase in young potentially fertile women may result in a comparable number of oocytes and comparable blastocyst euploidy rates compared with follicular phase stimulation. This may imply that in case of a freeze-all protocol in young patients with good ovarian reserve, clinicians may safely consider initiation of ovarian stimulation during the luteal phase. This research was supported by an unrestricted grant from MSD/Organon. N.P.P. has received Research grants and honoraria for lectures from: Merck Serono, MSD/Organon, Ferring Pharmaceuticals, Besins Intenational, Roche Diagnostics, IBSA, Theramex, Gedeon Richter. F.M., E.C., M.R. and S.G. declared no conflict of interests. The study was registered at Clinical Trials Gov (NCT03555942).

Intraovarian platelet-rich plasma administration could improve blastocyst euploidy rates in women undergoing in vitro fertilization.

ObjectivePlatelet-rich plasma (PRP) therapy has received a considerable attention as an adjunct to fertility treatments, especially in women with very low ovarian reserve and premature ovarian insufficiency. Although recent studies have demonstrated that PRP led to improvements in folliculogenesis and biomarkers of ovarian reserve, the effect of intraovarian PRP administration on embryo genetics has not been studied.MethodsWe report a pilot study of patients who had preimplantation genetic testing for aneuploidy (PGT-A) before and then within 3 months following PRP administration. Twelve infertile women with at least one prior failed in vitro fertilization (IVF) cycle underwent ovarian stimulation (cycle 1) with a gentle stimulation protocol and PGT-A performed at the blastocyst stage. Following cycle 1, autologous intraovarian PRP administration was performed. Within 3 months following PRP administration, the patients underwent cycle 2 and produced blastocysts for PGT-A. The percentage of euploid embryos between both cycles was compared.ResultsThe mean age of all participants was 40.08±1.46 years, and their mean body mass index was 26.18±1.18 kg/m2. The number of good-quality embryos formed at the blastocyst stage was similar between cycle 1 and cycle 2 (3.08±0.88 vs. 2.17±0.49, respectively; p=0.11). Among all patients in cycle 1, 3 of 37 embryos were euploid (8.11%) while in cycle 2, 11 out of 28 embryos were euploid (39.28%, p=0.002). Three clinical pregnancies were noted among this patient group.ConclusionThis novel study is the first to present an improvement in the embryo euploidy rate following intraovarian PRP application in infertile women with prior failed IVF cycles. The growth factors present in PRP may exhibit a local paracrine effect that could improve meiotic aberrations in human oocytes and thus improve euploidy rates. Whether PRP improves live birth rates and lowers miscarriage rates remains to be determined in large trials.

Relationship Between Number of Oocytes Retrieved and Embryo Euploidy Rate in Controlled Ovarian Stimulation Cycles

This cohort study is aimed to determine if higher number of oocytes retrieved affects the rate of euploidy in the embryos of women undergoing controlled ovarian stimulation (COS) for in vitro fertilization (IVF) with preimplantation genetic testing for aneuploidy (PGT-A). A negative trend between the number of oocytes retrieved and embryo euploidy rate was observed using Visual Analytics software, especially when a higher number of oocytes were retrieved. After regression analysis, patient age was the only variable found to have a statistically significant negative effect (p < 0.0001) on euploidy rate in all regression models. Number of oocytes retrieved was not found to have a statistically significant effect on euploidy rate when analyzed per number of biopsied blastocysts (p = 0.5356), per number of oocytes retrieved (p = 0.1025), and per number of fertilized oocytes (p = 0.7241). The parameter estimates in the linear regression models were negative for number of oocytes retrieved. This study shows a statistically significant effect between patient age and embryo euploidy rate, which is already known. There is some evidence to suggest that higher number of oocytes retrieved may negatively impact the number of euploid embryos per number of oocytes retrieved based on the visual analytic graphs, p value approaching significance, and the negative parameter estimates in the regression models.

P-539 Effects of gender of chromosomal translocation's carrier on pre-implantation genetic testing for structural rearrangement (PGT-SR) outcomes

Abstract Study question Do the outcomes of PGT-SR cycles vary according to the gender of chromosomal translocation’s carrier? Summary answer The rate of balanced embryos was significantly higher when translocation carrier was a male as compared to female, but clinical outcomes were not significantly different. What is known already It has been well described that chromosomal translocations lead to very high rate of unbalanced embryos, and have a subsequent negative impact on blastulation, implantation, miscarriage and live birth rates.PGT-SR allows selecting balanced embryos and increases the chances of pregnancy, while decreasing the risk of micarriage in couples with chromosomal translocations. However, whether the gender of translocation carrier is associated with significantly different impact on PGT-SR outcomes is not known. Study design, size, duration This is a monocentric retrospective cross-sectional study conducted in all PGT-SR cycles performed between 2015 and 2020. A total of 603 cycles, carried out in 283 couples, were included in the analysis. Participants/materials, setting, methods Two groups have been formed according to the gender of translocation carrier, Group 1 for female carriers (107 couples, 229 cycles), and group 2 for male (175 couples, 373 cycles). The rate of balanced embryos, ovarian stimulation parameters, number of oocytes, biopsy rate, clinical pregnancy rate and live birth rate were compared between both groups. Main results and the role of chance Female age and infertility etiologies were comparable between both groups. Mean total dose of gonadotrophin and peak E2 levels were not significantly different in both groups (2422± 895 units vs 2290 ± 858 respectively, p &amp;gt; 0.05) (2460± 1346 pg/ml vs 2497 ± 1397, p &amp;gt; 0.05). There was no significant difference in the number of oocytes and biopsied embryos rate between Group 1 and Group 2. The rate of balanced embryo after PGT-SR was significantly lower in group 1 (female carriers) than in group 2 (21.1% vs 24.8% respectively, p = 0.036). Finally, biochemical pregnancy rate and live birth rate per transfer were not statistically different between both groups (46.2% vs 43.2% and 20.3% vs 18.5% respectively, p &amp;gt; 0.05). Limitations, reasons for caution The retrospective nature of the study was its main limitation. PGT-A was not used, preventing from concluding on embryo euploidy rate. Wider implications of the findings Further fundamental studies should be performed to explore the respective mechanisms involved in chromosomal rearrangements according to gender. Our study suggests that a higher number of oocytes could be targeted in female translocation carriers in order to increase the probability of obtaining at least one balanced embryo. Trial registration number 2

Does Microfluidic Sperm Sorting Affect Embryo Euploidy Rates in Couples with High Sperm DNA Fragmentation?

Male infertility contributes as the main factor in 30-50% of infertility cases. Conventional methods for sperm preparation have induced questioning of sperm recovery rates. The microfluidic sperm sorting (MSS) technique selects highly motile sperm with lower levels of SDF (sperm DNA fragmentation) compared to conventional sperm sorting techniques. This study aimed to determine whether utilizing this technique will reveal better embryo quality and euploidy rates in couples with repeated implantation failure (RIF) and high SDF in a new PGT-A (preimplantation genetic testing for aneuploidies) cycle. This retrospective study included couples referred to PGT-A for previous repeated ART (assisted reproductive techniques) cycle failures and with high SDF. In their new cycles, couples who accepted the technique were assigned to the MSS group, and the rest were managed with DGC (density-gradient centrifugation). Two groups were compared in terms of fertilization and euploidy rates, clinical miscarriage and live birth rates, the total number of blastocysts, and top quality blastocysts. There was no difference between the groups regarding fertilization rates, euploidy rates, clinical miscarriage, and live birth rates. The total number of blastocysts and top quality blastocysts were significantly higher in the MSS group. The MSS technique provides a higher number of top-quality blastocysts than DGC; however, neither euploidy nor live birth rates improved. Studies focusing on confounding factors to embryonic genomic status in the presence of high SDF are needed.

Blastocyst euploidy rates in low-prognosis patients according to the POSEIDON criteria: a retrospective analysis of 3016 embryos

Research QuestionDo embryo euploidy rates differ in the four groups of women with low prognosis as stratified by the POSEIDON criteria? DesignThis was a retrospective cohort study of low-prognosis patients who met the POSEIDON criteria and underwent preimplantation genetic testing for aneuploidies (PGT-A) from January 2013 to June 2020 at the Center for Reproductive Medicine, Shandong University, China. A total of 3016 blastocysts from 1269 PGT-A cycles were included in the study. The primary outcome was the euploidy rate of the blastocysts. For each group, regression analyses were performed to quantitatively describe the relationship between maternal age and embryo euploidy rate. ResultsThe euploidy rate of embryos in women with poor ovarian response (POR) was 39.1% in total. There were 727, 1052, 275 and 962 blastocysts in groups 1, 2, 3 and 4, respectively, with corresponding embryo euploidy rates of 57.2%, 34.9%, 52.4% and 26.2% (P < 0.001). Within each group, the euploidy rate decreased with age, especially in women aged 35 years or older (i.e. groups 2 and 4). ConclusionsEuploidy rates were more favourable in groups 1 and 3, of a young age, re-emphasizing that oocyte quality is the primary factor determining embryo euploidy rate. The study's findings demonstrated the reasonability of categorizing women with POR by the POSEIDON criteria depending on female age and ovarian reserve biomarkers. These results also provide information for women with POR in different subgroups so they can receive proper counselling on the possible prognosis.

LASER-ASSISTED HATCHING ON DAY 1 DEMONSTRATES EQUIVALENT EMBRYO YIELD, TIMING OF BLASTOCYST DEVELOPMENT AND EMBRYO EUPLOIDY RATES COMPARED WITH STANDARD DAY 3 ASSISTED HATCHING

To compare in vitro fertilization (IVF) cycle outcomes including number of embryos cryopreserved, percentage of blastocysts cryopreserved on day 5 and embryo euploidy rate between cycles with assisted hatching (AH) on day 1 and day 3.

BLASTOCYST EUPLOIDY RATES ARE NOT AFFECTED BY INTRACYTOPLASMIC SPERM INJECTION (ICSI) VERSUS CONVENTIONAL INSEMINATION

To investigate whether embryo euploidy rates are affected by the method of oocyte insemination (ICSI vs. conventional insemination) within the same cohort of eggs. Single center retrospective cohort study. Only non-male factor patients were included. Patients’ eggs were divided into two groups after ova retrieval. In the first group, the eggs were stripped and mature eggs were fertilized by ICSI. In the second group, cumulus oocyte complexes were co-cultured with 200,000/ML sperm overnight (conventional insemination). Fertilization was checked between 16-18 hours after either ICSI or conventional insemination, and the fertilized embryos were cultured up to day 6. Trophectoderm biopsy and PGT-A analysis were performed at the blastocyst stage. The blastulation rates and euploidy rates were calculated for each patient using paired Student T-test. Data from a total number of 46 non-male factor patients were collected (mean age 36.6 years). There were 217 and 117 blastocysts biopsied in the ICSI and conventional insemination groups with an average blastulation rate of 54.4% vs. 53.3% (P=0.82), respectively. There were also no significant differences observed in the euploidy rates between ICSI and conventional insemination groups, respectively (49.2% vs. 45.2%, P=0.49). This study compared the effect of fertilization methods on euploidy rates using the same cohort of eggs. Results indicate that the blastulation and euploidy rates were not affected by the method of oocyte fertilization. Larger data set may be needed for further study.