Euploid Embryos Research Articles

The Association between Embryo Development and Chromosomal Results from PGT-A in Women of Advanced Age: A Prospective Cohort Study.

Embryo morphology and morphokinetics have been studied for their association with euploid embryos. However, the results are controversial, especially in the advanced-aged women group, when the risk of aneuploidy increases significantly. This prospective cohort study evaluated the association between embryo development between day-3 cleavage and day-5 blastocyst stages and euploidy rates, determined using preimplantation genetic testing for aneuploidy (PGT-A). Embryos from women aged 35 years and above who underwent intracytoplasmic sperm injections and PGT-A were studied. Day-3 cleavage-stage embryos were evaluated for their cell number, and day-5 blastocyst-stage embryos were evaluated for their morphological grade. Embryo development from day 3 to day 5 was categorized as either good or poor development and evaluated for its association with the PGT-A results. We evaluated 325 embryos from 101 infertile couples. It was found that 55.17% of blastocysts with good development and 29.83% with poor development were euploid. A significant association was found between embryo development and euploidy rates in advanced-aged women (p < 0.001). Also, there were significantly higher rates of euploid embryos with good blastocyst morphological grades, especially blastocyst expansion grades and trophectoderm grades. In conclusion, embryo morphokinetics shows promising results in predicting euploidy in advanced female age.

Sperm selection by zona adhesion improves assisted reproductive treatment outcomes.

Zona pellucida is one of the main selective barriers for the spermatozoa before reaching the oocyte. Using native zona in the sperm selection prior to Intracytoplasmic Sperm Injection (ICSI) has been proven effective but inconvenient approach in In Vitro Fertilisation (IVF) laboratory. The application of autologous solubilised zonae pellucidae in the sperm selection prior to ICSI has not been studied yet. To compare the assisted reproductive treatment (ART) outcomes (implantation, pregnancy, live birth, and miscarriage rates) after ICSI performed with spermatozoa selected on their ability to adhere to immobilised solubilised zonae pellucidae and conventionally selected spermatozoa. In total, 500 couples fulfilled the inclusion criteria and 368 of them were included in the study. After random allocation, 192 couples had spermatozoa selected by sperm-zona adhesion for ICSI (study group) and 176 patients underwent standard ICSI (control group). In the study group, patients' own zonae were acid solubilised and immobilised on petri dishes. The partner's motile spermatozoa were placed in the dishes and the adhered spermatozoa were used for ICSI. For the control group, the conventional sperm selection by morphological criteria was applied prior ICSI. All women underwent frozen ET with euploid embryos. Chi square test was used to compare the data. The sperm selection by zona adhesion resulted in significantly higher implantation rate (50.4%vs. 37.0%, p=0.003), clinical pregnancy rate (43.8%vs. 33.3%, p=0.018) and live birth rate (38.0%vs. 25.9%, p=0.004) and significantly lower incidence of miscarriage (11.3%vs. 22.2%, p=0.044) in comparison to the conventional method of the sperm selection. The application of solubilised zonae pellucidae in the sperm selection for ICSI benefits ART outcomes in couples with unexplained infertility. Moreover, sperm-zona selection significantly reduces the risk of miscarriages.

The morphokinetic signature of human blastocysts with mosaicism and the clinical outcomes following transfer of embryos with low-level mosaicism

BackgroundGenetic mosaicism is commonly observed in human blastocysts. Embryos’ morphokinetic feature observed from time-lapse monitoring (TLM) is helpful to predict the embryos’ ploidy status in a non-invasive way. However, morphokinetic research on mosaic embryos is extremely limited. Moreover, transfer of mosaic embryos is a new attempt in reproductive medicine, while studies regarding the clinical and neonatal outcomes following transfer of embryos with different levels and types of mosaicism are needed. This study aimed to investigate the morphokinetic characteristics of mosaic blastocysts, uncover clinical outcomes of mosaic embryos, and evaluate the effect of level and type of mosaicism on transfer outcomes.ResultsA total of 923 blastocysts from 229 preimplantation genetic testing cycles were cultured in TLM incubators in a single fertilization center between July 2016 and July 2021. Multivariate logistic regression models showed mosaic embryos had significantly shorter time to reach morula when compared with euploid (P = 0.002), mosaic with aneuploid (P = 0.005), and aneuploid (P = 0.005) embryos after adjusting the potential confounders. KIDScore is an artificial intelligence scoring program from time lapse incubation system to predict embryo implantation potential. Mosaic with aneuploid embryos had significantly lower KIDScore than euploid (P = 6.47e−4), mosaic (P = 0.005), and aneuploid (P = 0.004) embryos after adjustment. Meanwhile, we compared the clinical outcomes following transfer of low-level (< 50%) mosaic embryos (N = 60) with euploid embryos (N = 1301) matched using propensity scoring collected from September 2020 to January 2023. Mosaic embryos had significantly lower clinical pregnancy rate (41.67% vs. 57.65%, P = 0.015) and live birth rate (38.33% vs. 51.35%, P = 0.048) than the euploid embryos. Subgroup analyses showed the whole, segmental, and complex chromosome mosaic embryos had the similar clinical outcomes.ConclusionsThe shortened time to reach morula in mosaic embryos and the low KIDScore in mosaic with aneuploid embryos revealed innovative clues to embryo selection with the non-invasive TLM and provided new insights into biological mechanism of chromosomal abnormality. The analyses of overall and subgroups of mosaic embryo transfer outcomes helped to optimize embryo transfer scheme for in-vitro fertilization procedures. Multi-center prospective studies with large sample sizes are warranted to validate our results in the future.

Beyond appearance: Can morphologically low-grade euploid blastocysts yield successful pregnancies?

The primary objective of this investigation is to evaluate how morphological quality affects the pregnancy outcomes in euploid embryos determined by preimplantation genetic testing for aneuploidies (PGT-A). Concurrently, as a secondary objective, we aim to identify which specific aspects of morphological evaluation exert the most significant impact on these outcomes. A retrospective analysis of 451 single euploid embryo transfer cycles at our clinic was conducted. Embryos were evaluated based on the degree of blastocyst expansion, inner cell mass (ICM), trophectoderm (TE) morphology, and the day of blastocyst vitrification. Outcomes between morphologically low-grade and high-grade embryos were compared. Additionally, the study analyzed which morphological factors most influenced pregnancy outcomes. Pregnancy outcomes were significantly lower in morphologically low-grade blastocysts compared to high-grade ones. Among the morphological evaluations, the ICM assessment was significantly associated with the live birth rate. Our study indicates that the morphological quality of euploid embryos, particularly the evaluation of the ICM, plays a crucial role in IVF-ET success.

Preimplantation genetic testing for complex chromosomal rearrangements: clinical outcomes and potential risk factors.

Complex chromosome rearrangements (CCR) are rare structural abnormalities involving at least three breakpoints, categorized into three types based on their structure: type A (three-way rearrangements), type B (double two-way translocations), and type C (exceptional CCR). However, thus far, limited data exists on preimplantation genetic testing for chromosomal structural rearrangements (PGT-SR) in CCR carriers. This study aims to evaluate the clinical outcomes and influencing factors of PGT-SR in couples with CCR. Fifteen couples with unique CCR recruited from 793 couples following PGT-SR between January 2017 and May 2023. In addition, a total of 54 CCR cases, 39 previously reported as well as 15 newly added, were included in the analysis of factors associate with normal/balanced embryos. A total of 100 blastocysts were biopsied and analyzed in 15 CCR couples after 17 PGT-SR cycles, with 16.0% being euploid, 78.0% aneuploid and 6.0% mosaic. 11 normal/balanced embryos and one mosaic embryo were transferred, resulting in eight live births. Furthermore, based on the combined data from 54 CCR carriers, the proportion of normal/balanced embryos was 10.8%, with a significant decrease observed among female carriers compared to male heterozygotes (6.5% vs. 15.5%, p = 0.002). Type B exhibited the lowest rate of euploid embryos at only 6.7%, followed by type A at 11.6% and type C at 14.0%, although the differences were not significant (p = 0.182). After completing the multivariate generalized estimating equation (GEE) analysis, type B (p = 0.014) and female carrier (p = 0.002) were identified as independent risk factors for fewer euploid embryos. The occurrence of balanced CCR in patients with reproductive abnormalities may be more frequent than we expected. Despite the proportion of normal/balanced embryos being significantly low, which can be influenced by CCR type and carrier's sex, PGT-SR may improve the reproductive outcomes among CCR cases. These findings can optimize the clinical management and genetic counseling of CCR carriers seeking assisted reproductive technology (ART).

Management strategies following implantation failure of euploid embryos.

Euploid blastocyst implantation failure may result from embryonic factors undetectable by preimplantation genetic testing for aneuploidy (PGT-A); however, various nonembryonic factors can also intricately interfere with implantation. This review seeks to clarify evidence-based testing and treatments for implantation failure after euploid embryo transfer. We conducted a review of the literature on implantation failure after euploid embryo transfer or multiple embryo transfer cycles, which mainly included systematic reviews and meta-analyses. The recommended tests for implantation failure include (1) hysteroscopy, (2) endometrial CD138 immunohistochemistry and bacterial culture, (3) serum 25-hydroxyvitamin D3, and (4) thrombophilia screening. Based on diagnostic findings, the following treatments have been recommended: (1) antibiotics for chronic endometritis, (2) vitamin D replacement, (3) lifestyle modification, and (4) low-dose aspirin starting from the postimplantation period for thrombophilia. Moreover, frozen-thawed single euploid blastocyst transfer using assisted hatching and hyaluronan-enriched transfer medium may support embryo implantation. To ensure a successful pregnancy in subsequent embryo transfers, simple, inexpensive, and evidence-based tests and treatments should be selected.

The use of combined hormonal contraceptive for in-vitro fertilization cycle priming is not associated with decreased pregnancy rate in frozen embryo transfer cycles: a cohort study.

The objectives of this study are to evaluate the cycle outcomes from IVF treatment preceded by oral contraceptive pills (OCP) priming compared to estradiol pretreatment and to determine if there is a role for OCP priming for those undergoing frozen embryo transfers. The study took place at a university-affiliated fertility center in Canada. The study included in-vitro fertilization (IVF) antagonist cycles from Jan 2016 to Jun 2019. Those with protocol deviation or treatment cancellation were excluded. There were 2237 cycles by 1958 patients; 27% of cycles utilized OCP priming. The average age in the OCP group was 34 years old compared to 36.5 in the estradiol group (P<0.01). AMH was reported in 43% of patients and was 3.7ng/mL in the OCP group versus 2.2 ng/mL in the estradiol group (P<0.01). The number of oocytes (15.2 vs. 12.5) and number of blastocysts (4.6 vs. 3.3) were higher in the OCP group (P all <0.01). After adjusting for age and AMH with linear regression for the 978 cycles with recorded AMH (24% with OCP prime), a significantly higher number of oocytes (13.8 vs. 11.9, P=0.002) was still noted in the OCP group. There were 866 euploid embryo transfer cycles (28% with OCP prime). There were no significant differences in implantation (77% vs. 76%) or ongoing pregnancy rates (56% vs. 54%) between those who had a frozen embryo transfer after OCP primed compared to estradiol primed stimulation cycles (P all >0.6). There were no differences in pregnancy outcomes from euploid frozen blastocyst transfers after OCP primed antagonist cycles compared to estradiol pretreatment. In fact, the use of OCP pretreatment was associated with increased oocyte yield, keeping in mind demographic differences with the OCP pretreatment group being younger with higher anti-Müllerian hormone and a higher prevalence of PCOS. Thus, OCP priming should still be considered in specific populations, such as those with oligo-ovulation or adequate ovarian reserve.

Differential Gene Regulation of the Human Blastocyst Trophectoderm and Inner Cell Mass by Progesterone.

Knowledge of action of progesterone (P4) on the human preimplantation embryo is lacking. The objective of this study was to determine expression of a mitochondrial P4 receptor (PR-M) in the trophectoderm (TE) and the inner cell mass (ICM) of the human blastocyst and to determine P4-induced gene expression during growth from the cleavage to the blastocyst stage. Previously cryopreserved cleavage stage embryos were treated with P4 (10-6 M) or vehicle until blastocyst development. Cells from the TE and the ICM of dissected euploid embryos underwent RNA-seq analysis, while other embryos were used for analysis of nuclear PR (nPR) and PR-M expression.PR-M expression was confirmed in the TE, the ICM, and a human embryonic stem cell line (HESC). Conversely, nPR expression was absent in the TE and the ICM with low expression in the HESC line. RNA-seq analysis revealed P4 effects greater in the TE with 183 significant pathway changes compared to 27 in the ICM. The TE response included significant upregulation of genes associated with DNA replication, cell cycle phase transition and others, exemplified by a 7.6-fold increase in the cell proliferation gene, F-Box Associated Domain Containing. The majority of ICM pathways were downregulated including chromosome separation, centromere complex assembly and chromatin remodeling at centromere. This study confirms that human blastocysts express PR-M in both the TE and the ICM, but lack expression of nPR. P4-induced gene regulation differs greatly in the two cell fractions with the predominant effect of cell proliferation in the TE and not the ICM.

Identification of cryptic balanced translocations in couples with unexplained recurrent pregnancy loss based upon embryonic PGT-A results.

The goal of this study is to determine whether any balanced translocation (BT) had been missed by previous karyotyping in patients with unexplained recurrent pregnancy loss (uRPL). This case series included 48 uRPL-affected couples with normal karyotypes. The embryos from these couples have all undergone preimplantationtesting for aneuploidies (PGT-A). Based on the PGT-A's results, 48 couples could be categorized into two groups: 17 couples whose multiple embryos were detected with similar structural variations (SVs, segmental/complete) and 31 couples without such findings but who did not develop any euploid embryo despite at least three high-quality blastocysts being tested. The peripheral blood sample of each partner was then collected for mate-pair sequencing (MPseq) to determine whether any of them were BT carriers. MPseq analyses identified 13 BTs in the 17 couples whose multiple embryos had similar SVs detected (13/17, 76.47%) and three BTs in the 31 couples without euploid embryo obtained (3/31, 9.7%). Among the 16 MPseq-identified BTs, six were missed due to the limited resolution of G-bandingkaryotyping analysis, and the rest were mostly owing to thesimilar banding patterns and/or comparable sizes shared by the two segments exchanged. A normal karyotype does not eliminate the possibility of carrying BT for couples with uRPL. The use of PGT-A allows us to perceive the "carrier couples" missed by karyotyping analysis, providing an increased risk of finding cryptic BTs if similar SVs are always detected on two chromosomes among multiple embryos. Nonetheless, certain carriers with translocated segments of sub-resolution may still go unnoticed.

Y chromosome AZFc microdeletion may have negative effect on embryo euploidy: a retrospective cohort study

BackgroundEmbryo aneuploidy is a main of principal reason of pregnancy loss, in vitro fertilization (IVF) failure and birth defects in offspring. Previous researchs have demonstrated that Y chromosome AZFc microdeletion was associated with reproduction outcomes, however, the relationship between Y chromosome AZFc microdeletion and embryo aneuploidy remains unexplored.MethodsThis retrospective cohort study enrolled 513 patients with 603 cycles in the reproductive center of Nanjing Maternity and Child Health Care Hospital from January 1, 2016 to June 30, 2022. The study cohort was divided into two groups: the AZFc microdeletion group, comprising 53 patients and 58 cycles, and the control group, comprising 460 patients and 545 cycles. Statistical methods including restricted cubic spline and generalized estimating equation (GEE) were employed to evaluate the relationship between Y chromosome AZFc microdeletion and embryo euploidy.Results294 and 2833 blastocysts were selected as AZFc microdeletion group and control group, respectively. Patients with Y chromosome AZFc microdeletion had significantly higher embryo aneuploid rate (33.0% vs. 27.3%, P < 0.05), lower rate of normal fertilization rate (81.5% vs. 90.3%, P < 0.05) and lower blastocysts formation rate (47.0% vs. 57.8%, P < 0.05) compared with the control group. However, no significant differences in pregnancy outcomes after euploid embryos transfer were observed between these two groups.ConclusionsOur study underscored the association between Y chromosome AZFc microdeletion and an elevated risk of embryo aneuploidy. Before the conventional intracytoplasmic sperm injection (ICSI) treatment, couples with Y chromosome AZFc microdeletion should be apprised of the heightened susceptibility to embryo aneuploidy. Preimplantation genetic testing for aneuploidy (PGT-A) should be introduced for selection.

#190 : Impact of Fertilisation Techniques and Day of Biopsy on Pre-Implantation Genetic Testing Outcomes: A Retrospective Study

Background and Aims: Pre-implantation genetic testing (PGT) is a technology utilised in assisted reproductive technologies (ART) to aid in embryo selection and reduce time to pregnancy. However, the impact on PGT outcomes of different fertilisation techniques, to date, has been the subject of a limited number of studies. Therefore, we investigated the effect of three different fertilisation methods - intracytoplasmic sperm injection (ICSI), PIEZO-ICSI, and standard in vitro fertilisation (IVF) - on PGT outcomes, with further analysis performed for day of biopsy. Method: Retrospective analysis of 760 embryos biopsied for PGT from 290 patients undergoing IVF treatment in 2021. Fertilisation was achieved via ICSI, PIEZO-ICSI, or IVF, followed by embryo culture using a sequential media system. Trophectoderm biopsy samples were analysed using a Next Generation Sequencing based assay (Illumina VeriSeq PGS). Ploidy outcomes from PGT (euploid, aneuploid, mosaic, inconclusive and no result) were interrogated for differences between fertilisation method or day of biopsy (Day 5 or 6). For analyses, frequency data of ploidy outcomes were reported as percentages and divided into two maternal age groups: &lt;38 and [Formula: see text]38 years. Statistical differences were determined using a chi-squared test. Results: Fertilisation technique did not have a significant effect on ploidy across both maternal age cohorts (% euploid embryos: &gt;38 years old (n=431) PIEZO-ICSI 61% vs ICSI 58% vs IVF 51%; p&gt;0.05, and [Formula: see text]38 years old (n=329) PIEZO-ICSI 37% vs ICSI 40% vs IVF 39%; p&gt;0.05). For both age groups, no significant differences were observed in PGT outcomes for Day 5 biopsy compared to Day 6 (% euploid embryos: &lt;38 years old (n=431) Day 5 62% vs Day 6 54%; p&gt;0.05, and [Formula: see text]38 years old (n=329) Day 5 45% vs Day 6 37%; p&gt;0.05). Conclusion: The study suggests that different fertilisation techniques or the day of biopsy may not influence PGT outcomes.

#155 : Embryo Thaw and Biopsy - A Clinical Guide for Patients

Background and Aims: Preimplantation genetic testing allows the selection of chromosomally normal embryos for transfer, and therefore may reduce the time required to live birth. Some patients request to perform biopsy on previously cryopreserved embryos. This includes situations where the embryos were frozen untested, or no result was obtained from the initial trophectoderm biopsy. Method: We analysed the outcomes of 108 embryos over 3 years in the effort to provide guidance to patients in terms of the expectations when considering performing thaw and biopsy on cryopreserved embryos. This includes 32 untested embryos (planned for PGT) (Group A), 45 embryos frozen without the intention of testing (Group B) and 34 embryos that had no results obtained in the first biopsy (Group C). The developmental timing of the embryos in the initial culture and the extended culture timing following thawing, was examined using timelapse footage. Results: Among the 108 embryos that were thawed and subsequently cultured, 78.7% embryos were suitable for biopsy with an average of 8.5 h in extended culture. Group A had the lowest proportion of embryos suitable to biopsy after extended culture (56.3%), which is significantly lower than Group B (83.3%, P=0.0181) and Group C (94.1%, P=0.0004). Interestingly, Group A had the lowest proportion of euploid embryos (16.7%), followed by Group B (40.0%) and Group C (46.9%). Interestingly, of the Group C embryos that previously failed to amplify only 26.7% were euploid, compared to 64.7% for those that previously amplified but failed to pass quality control. There was no correlation between time to blastulate, time to form full blastocyst or culture time and euploidy rate. At the time of our study, 18 of the 32 euploid embryos were thawed and transferred, resulting in 44.4% clinical pregnancy rate. Conclusion: This analysis provides important information to guide patients’ expectations at a clinical level.

Embryonic, genetic and clinical outcomes of fresh versus vitrified oocyte: A retrospective cohort study

Objective: To compare embryonic development, ploidy status and clinical outcomes between fresh and frozen-thawed oocytes. Methods: This retrospective cohort study evaluated 83 fertilization cycles including both fresh and frozen oocytes from 79 patients at the HP Fertility Center of Hai Phong International Hospital of Obstetrics and Pediatrics in Vietnam. The patient underwent several ovarian stimulation cycles to accumulate a certain number of oocytes that would be vitrified. In the last oocyte retrieval, all patient's oocytes including both frozen and fresh would be fertilized. The outcomes included the rates of oocyte survival, cleavage embryo, blastocyst, ploidy status, pregnancy, biochemical pregnancy and clinical pregnancy. Results: The oocyte survival rate after thawing was 96.5%. No statistically significant difference was found when comparing fresh and frozen oocytes regarding fertilization rate (78.1% vs. 75.5%, P=0.461), usable cleavage embryo rate (86.9% vs. 87.2%, P=0.916) but usable blastocyst rate was found higher statistically in the frozen oocyte group (44.4% vs. 54.0%, P=0.049). The percentages of euploid, aneuploid and mosaic embryos between the fresh group and the vitrified group had no significant differences (33.8% vs. 31.6%, P=0.682; 51.0% vs. 54.2%, P=0.569; 15.2% vs. 12.4%, P=0.787; respectively). The rates of pregnancy, biochemical pregnancy and clinical pregnancy had no statistical difference (68.8% vs. 64.8%, P=0.764; 12.5% vs. 3.6%, P=0.258; 37.5% vs. 46.4%, P=0.565). 17 Mature oocytes are the minimum to have at least one euploid embryo. Conclusions: Oocyte vitrification does not affect embryonic, genetic and clinical results. The number of mature oocytes should be considered for fertilization in some cases.

#144 : Randomized Controlled Trial Evaluating Efficacy of Autologous Platelet-Rich Plasma Therapy for Patients with Recurrent Implantation Failure in Frozen-Thawed Embryo Transfer After PGT-A

Background and Aims: Despite keen development of ART, recurrent implantation failure (RIF) remains a challenging dilemma for fertility specialists. As it contains significant growth factors involved in delicate process of implantation, platelet–rich plasma (PRP) therapy should promote endometrial receptivity and improve ART. Objective: To evaluate if the intrauterine perfusion with autologous PRP enhances frozen-thawed embryo transfer effectiveness in patients with RIF after PGT-A. Methods: Study type: Interventional. Design: randomized controlled study Intervention Model: Parallel Assignment Masking: open label After obtaining institutional review board approval, 232 women aged 28-42 years were involved. Matching criteria: RIF, normal karyotype, absence of uterine factors of infertility, availability of euploid embryos after PGT-A (NGS). 2 groups of patients: study group (N = 118): single IP with 2.0 ml of autologous PRP on day 10-11 of menstrual cycle; Control group: no therapy (N = 114). Endometrium preparation was carried out according to standardized protocol. Primary outcome measure was clinical pregnancy rate. Secondary outcome measures were pregnancy loss rate, endometrial thickness and adverse event. Results: The clinical pregnancy rate was higher in the study group (63.55% vs 38.59%) ([Formula: see text]2=14.462, OR=2.775, 95% CI 1.630 - 4.722, p&lt;0,001). The endometrium thickness before intervention didn’t between groups (7.3 vs 7.4 mm), however, endometrium thickness measured just before embryo transfer was significantly higher in the study group (10.5 vs 8.4 mm, Student’s t-test value: 11.87; number of degrees of freedom f=230; Critical value of Student’s t-test=1.972, at significance level [Formula: see text]= 0.05). The pregnancy rate loss did not differ between groups ([Formula: see text]2=0.033, OR=0.908, 95% CI 0.324 – 2.546, p&gt;0,05). No adverse event was noted. Conclusions : – The PRP intrauterine perfusion should be considered perspective, safe and cost-effective therapy method for patients with RIF. – PRP does not influence on pregnancy loss rate – Meta-analysis is required. Support: None

#121 : The Easier Blastocyst Biopsy, The More Trophectoderm Cells We Got

Background and Aims: Several studies showed various factors associated with pregnancy outcomes after the euploid embryo transfer, including parental, endometrial, and embryo biopsy. The consensus of the most suitable biopsied cell number is 5-10 cells among IVF labs. Therefore, for embryologists, how to have a steadily manipulated blastocyst to get an appropriate biopsied trophectoderm cell number is crucial. However, the factors related to biopsy manipulation are still being determined. Here, we retrospectively analyze the DNA concentrations from biopsied cells to test the hypothesis TE grading, hatching status, and biopsy day correlated with embryologists’ biopsy handling. Method: A retrospective study enrolled 212 blastocysts from 49 PGT-A cycles (NGS platform) from September 2021 and March 2023 at the Fertility and Reproductive Genetic Center of Linkou Chang Gung Memorial Hospital, Taoyuan, Taiwan. All the TE cells were removed by mechanical cutting by embryologists following the lab standard operating procedure. This study excluded five blastocysts (2.4%, 5/212) for the unsuccessful whole genome amplification (WGA), i.e., DNA concentration was less than 10 ng/[Formula: see text] l. Results: No difference in average DNA concentration was found between embryo ploidy status (Fig. A, P=0.8), embryologist (Fig. A, P=0.65), and TE grading (Gardner TE grading B vs. C, P=0.8). However, combined hatching status and day of the biopsy revealed a significantly higher DNA concentration in the hatching Day 5 to Day 7 groups than in the hatched Day 5 to Day 7 group (Fig. B, P[Formula: see text]0.0001). Conclusion: Our data suggest that the embryo ploidy status and Gardner grading of TE cells did not affect embryologists’ steadily performing biopsies. Interestingly, the easier biopsy, the more TE cells we got. This result implies that the hatching (Gardner expansion grade 5) embryo has a higher potential for damage after trophectoderm biopsy.

#89 : Higher Euploidy Rate in Embryos Generated by ICSI Using Sperm Obtained After Microfluidics Sperm Sorting (MFSS). A Randomised Controlled Trial

Background and Aims: MFSS is a state-of-the-art technology that has the ability to sort a subpopulation of sperm cells with lower DNA fragmentation index (DFI), higher motility and higher normal forms without the need for centrifugation. Methods: A randomised controlled trial that included 925 couples that were planned for ICSI with day 5/6 biopsy, PGT-A with Next Generation Sequencing (NGS) and blastocyst vitrification. The embryo transfer was planned for future cycles. The inclusion criteria for women were age (18–45), BMI (18–30), AMH level more than 1.1 ng/ml, regular menstrual cycle and normal uterine ultrasound. Whereas the inclusion criteria for men were age (18–50), fresh semen sample with count more than 10 million/ml and total motility more than 30%. Semen samples were randomly assigned with 1:1 ratio to either preparation by routine density gradient (DG) or (MFSS). The primary outcome of the study is PGT-A aneuploidy results. The secondary outcome is the fertilisation rate, and blastocyst development rate. Results: The study was performed between January 2020 and July 2021. 463 semen samples were prepared with density gradient and 462 semen samples were sorted with microfluidic sperm sorting. All of the prepared semen samples were used for ICSI. The euploid embryo rate for the embryos resulted from density gradient preparation was 53.1% versus 59.3% for the embryos resulted from microfluidics sperm sorting (p=0.0034). The fertilisation rate was 77.9% for density gradient group versus 78.3% microfluidics group (p=0.063) and blastocyst rate was 51.6% for the density gradient group versus 59.2% for the microfluidics group (p=0.058). Conclusion: Semen samples prepared with MFSS for ICSI generated higher euploidy embryos following PGT-A with NGS.

#248 : Maternal Hypercholesterolemia Would Increase the Incidence of Embryo Aneuploidy in Couples with Recurrent Implantation Failure

Background and Aims: The association of dyslipidemia with reproductive outcomes is largely unknown, especially in recurrent implantation failure (RIF) patients. This study aimed to explore the impact of abnormal blood lipid levels on embryo genetic status and pregnancy outcomes in unexplained RIF (uRIF) patients after preimplantation genetic testing for aneuploidy (PGT-A). Method: In this retrospective study, they were divided into 4 groups according to the levels of cholesterol and triglyceride: non-hyperlipidemia group (NonH group), simple hypercholesterolemia group (SHC group), simple hypertriglyceridemia group (SHC group) and mixed hyperlipidemia group (MixH group). Additionally, patients were divided into 2 groups according to their HDL-C level. Embryos’ genetic status and pregnancy outcomes after transfer of euploid embryos were analyzed. Binary logistic regression and/or generalized estimating equation model were conducted to investigate the association of different types of dyslipidemia with aneuploid embryo rate and cumulative live-birth rate. Results: A total of 474 women were divided into four groups: NonH group (N=349), SHC group (N=55), SHT group (N=52) and MixH group (N=18). Compared with the NonH group, SHC group had a significantly increased aneuploid embryo rate [48.3% vs. 36.7%, P=0.006; adjusted OR (95% CI) = 1.52(1.04-2.22)], as well as a reduced number of good-quality blastocysts [3.00±2.29 vs. 3.74±2.77, P=0.033]. The SHC group showed a lower cumulative live-birth rate (47.0% vs. 40.0%), good birth outcome (37.2% vs. 34.5%) and a higher risk of clinical pregnancy loss (11.1% vs. 17.9%) but did not reach statistical significance. The incidences of obstetric or neonatal complications and other adverse events were similar in the four groups. Conclusion: We found that uRIF women with hypercholesterolemia had an increased proportion of aneuploid embryos and a reduced number of high-quality embryos, while different types of hyperlipidemias had no correlation with cumulative live birth rate as well as pregnancy and neonatal outcomes.

Can We Really Make a Difference in Reducing Miscarriage?

Many miscarriages occur due to chromosomal problems, and may not be preventable. But there are many miscarriages that result from the loss of euploid embryos. Why are these pregnancies lost? Where is the problem, and what can be done? There is a pervasive and harmful idea that miscarriages are nature’s way of dealing with an abnormal pregnancy, and therefore they cannot really be prevented, and we do not need to try to prevent them. This idea ignores the fact that there are several steps that can be taken to reduce the risk of miscarriage. This lecture asks the questions: what can a woman or couple do to reduce their risk of miscarriage? What can a healthcare provider do to reduce the risk of miscarriage? What is evidence? What are the limitations in the evidence? The recently updated evidence-based NICE, RCOG and ESHRE guidelines form the foundation of this lecture.

#279 : The Relationship Between Maternal Age and the Chromosomal Status of the Embryo

Background and Aims: Maternal age is one of the success factors of in vitro fertilization (IVF). Specifically, advanced maternal age (AMA; defined as [Formula: see text]35 years) shows just a negligible impact upon fertilization rate and a mild impact upon embryo development to the blastocyst stage, but results in a dramatic impact upon blastocyst aneuploidy rate. This is also associated with a progressive decrease in ovarian stimulation so that the number of oocytes and embryos produced from an older maternal age becomes less and the implantation rate decreases. However, the high incidence of embryonic chromosomal abnormalities in young women does not exclusively prove that maternal age is related to the quality of the resulting embryos. This study aims to analyze the relationship between maternal age and the chromosomal status of the embryo. Method: This cross-sectional research was done on 30 embryos from 12 patients aged 28-40 years old who underwent an IVF program in three clinics. Embryos were cultured until blastocyst stage. Preimplantation genetic testing for aneuploidy (PGT-A) was done after blastocyst biopsy on day 5 or 6, and chromosomal analysis was done using the next generation testing (NGS) method. Then the relationship between maternal age with embryo chromosomal status was analysed. Results: There is no difference in subjects’ characteristics between the euploid and aneuploid embryo groups (p&gt;0.05). There is no significant difference in maternal age of the euploid and aneuploid embryo groups. (With p=0.690). Conclusion: Maternal age has no relationship with embryo chromosomal status. There is no specific pattern in the relationship between age and chromosomal status, except in the age range of 36 to 40 years, it is found that increasing age increases the number of aneuploid embryos and decreases the number of euploid embryos.

#361 : What Does My Embryo Score Mean? The Relationship Between KIDScore, Life Whisperer AI Score, and Embryo Euploidy

Background and Aims: Many models have been developed for predicting an embryo’s euploidy or implantation potential. These models assign continuous scores to embryos, based on annotation performed by embryologists according to morphokinetic parameters, or based on deep learning analysis of embryo images or video, with higher scores indicating higher probability of implantation and/or euploidy. However, the actual probability of implantation or euploidy at a given score is not always clear. We aimed to investigate the relationship between scores assigned by an annotation model (KIDScoreD5 v3.1, Vitrolife), an image analysis model (Life Whisperer, Presagen), and embryo euploidy. Method: From January 2019 to May 2021, PGT-A by NGS performed on blastocysts in Thomson Hospital Kota Damansara in Malaysia identified 273 euploid and 212 aneuploid blastocysts. Since blastocyst culture in a timelapse incubator (Embryoscope+, Vitrolife) is routine in our centre, the KIDScore for each blastocyst was known. Still image data of the same embryos was uploaded to the Life Whisperer (LW) platform. Embryos were grouped based on their KIDScore and LW score, and the percentage of euploid and aneuploid embryos in each group was calculated. Results: In both models, there were more embryos in higher score groups, as only embryos with good morphology underwent PGT-A (Figures 1 and 2). In KIDScore groups up to score 8, approximately 50% of embryos were euploid, while in KIDScore groups &gt;8-9 and &gt;9-10, 76.6% and 75.5% of embryos were euploid respectively (Figure 1). In LW groups up to score 7, approximately 50% of embryos were euploid, while in LW groups &gt;7-8, &gt;8-9, and &gt;9-10, the percentage increased to 56.1%, 58.6%, and 66.7% respectively (Figure 2). Conclusion: Increasing KIDScores and LW scores correspond with increased percentages of euploid embryos. However, the increase is not constant, especially for KIDScores. These results can be used to estimate the probability of euploidy at a given embryo score.