Blastulation Rate Research Articles

Experience in using biosimilars in IVF programs with donor oocytes in protocols with micronized progesterone

Relevance: Stimulation of oocyte donors' superovulation occupies a key place in the development of general assisted reproduction technology (ART). Oocyte donation programs involve young and healthy women undergoing severe ovarian stimulation protocols to obtain good-quality oocytes. Advances in recombinant DNA technology led to the development of recombinant follitropin alfa.The study aimed to evaluate the effectiveness of the biosimilar folitropin alfa in the superovulation stimulation cycles of oocyte donors in comparison with the original medication in Progestin primed ovarian stimulation (PPOS) protocols.Materials and Methods: A prospective cohort study included 25 oocyte donors who underwent superovulation induction. The first group consisted of 25 oocyte donors in whom superovulation was stimulated with a bio-analogue of follitropin alfa. The second group (control) included the same 25 oocyte donors stimulated with the original rFSH medication. The donors of the leading group were stimulated with biosimilar recombinant follicle-stimulating hormone and human menopausal gonadotropin. For the control group, a protocol was used using the original drug rFSH and CHMG in the same doses. The obtained donor oocytes were fertilized with ICSI or frozen. The program's result was determined by taking a blood test for b-hCG 14 days after embryo transfer.Results: The number of mature oocytes in the study groups did not differ significantly, amounting to 20.6 ± 1.1 in group 1 and 21.2 ± 1.3 in group 2. The average number of fertilized oocytes in group 1 was 5.6 ± 0.8, and in group 2 – 6.1 ± 1.1; thus, the fertilization rate in the two groups did not differ (67,5% in group 1 vs 79,2% in group 2, р> 0,05). The blastulation rate (49.1% in group 1 and 52.2% in group 2) did not differ significantly in both the biosimilar group and the original medication. The Top Quality Blastocysts in Group 1 were 1.5 ± 0.7 (55%) vs 1.8 ± 0.5 (48%) in Group 2. At the same time, the level of NNB in both groups also had no statistically significant differences.Conclusion: Sinnal-F was not inferior to the original medication in terms of the proportion of mature oocytes in oocyte donors. The study's results confirm that Synnal-f demonstrates favorable and comparable efficacy to the original medication in oocyte maturation in IVF/ICSI cycles.

Abnormal cleavage up to Day 3 does not compromise live birth and neonatal outcomes of embryos that have achieved full blastulation: a retrospective cohort study.

Do embryos displaying abnormal cleavage (ABNCL) up to Day 3 have compromised live birth rates and neonatal outcomes if full blastulation has been achieved prior to transfer? ABNCL is associated with reduced full blastulation rates but does not impact live birth rates and neonatal outcomes once full blastulation has been achieved. It is widely accepted that ABNCL is associated with reduced implantation rates of embryos when transferred at the cleavage stage. However, evidence is scarce in the literature reporting birth outcomes from blastocysts arising from ABNCL embryos, likely because they are ranked low priority for transfer. This retrospective cohort study included 1562 consecutive autologous in vitro fertilization cycles (maternal age 35.1 ± 4.7 years) performed at Fertility North, Australia between January 2017 and June 2022. Fresh transfers were performed on Day 3 or 5, with remaining embryos cultured up to Day 6 before vitrification. A total of 6019 embryos were subject to blastocyst culture, and a subset of 664 resulting frozen blastocysts was included for live birth and neonatal outcome analyses following single transfers. ABNCL events were annotated from the first mitotic division up to Day 3, including direct cleavage (DC), reverse cleavage (RC) and <6 intercellular contact points at the 4-cell stage (<6ICCP). For DC and RC in combination, the ratios of affected blastomeres over the total number of all blastomeres up to Day 3 were also recorded. All pregnancies were followed up until birth with gestational age, birthweight, and sex of the baby being recorded. Full blastulation rates for embryos showing DC (19.5%), RC (41.7%), <6ICCP (58.8%), and mixed (≥2) ABNCL types (26.4%) were lower than the rates for those without ABNCL (67.2%, P < 0.01 respectively). Subgroup analysis showed declining full blastulation rates with increasing ratios of combined DC/RC affected blastomeres over all blastomeres up to the 8-cell stage (66.2% when 0 affected, 47.0% when 0.25 affected, 27.4% when 0.5 affected, 14.5% when 0.75 affected, and 7.7% when all affected, P < 0.01). However, once full blastulation had been achieved, no difference was detected between DC, RC, <6ICCP, and no ABNCL blastocysts following single frozen transfers in subsequent live birth rates (25.9%, 33.0%, 36.0% versus 30.8%, P > 0.05, respectively), gestational age (38.7 ± 1.6, 38.5 ± 1.2, 38.3 ± 3.5 versus 38.5 ± 1.8 weeks, P > 0.05, respectively) and birthweight (3343.0 ± 649.1, 3378.2 ± 538.4, 3352.6 ± 841.3 versus 3313.9 ± 509.6 g, P > 0.05, respectively). Multiple regression (logistic or linear as appropriate) confirmed no differences in all of the above measures after accounting for potential confounders. Our study is limited by its retrospective nature, making it impossible to control every known or unknown confounder. Embryos in our dataset, being surplus after selection for fresh transfer, may not represent the general embryo population. Our findings highlight the incremental impact of ABNCL, depending on the ratio of affected blastomeres up to Day 3, on subsequent full blastulation. The reassuring live birth and neonatal outcomes of ABNCL blastocysts imply a potential self-correction mechanism among those embryos reaching the blastocyst stage, which provides valuable guidance for clinical practice and patient counseling. This research is supported by an Australian Government Research Training Program (RTP) Scholarship. All authors report no conflict of interest. N/A.

Developmental potential of non- and mono-pronuclear zygotes and associated clinical outcomes in IVF cycles.

This study aims to evaluate the developmental potential of 0PN, 1PN, and 2PN zygotes in IVF cycles and compare their clinical outcomes. We conducted a retrospective cohort study involving IVF patients. Blastocyst formation rates were assessed with 0PN, 1PN, and 2PN zygotes. Subsequently, we collected clinical outcome data following the transfer of these zygotes. The overall blastulation rate was similar between 0PN (29.6%) and 2PN (32.1%) zygotes, but 1PN zygotes exhibited a significantly lower blastulation rate (17.0%) compared to both 0PN and 2PN zygotes. Similarly, the overall rate of good-quality blastulation was comparable between 0PN (15.3%) and 2PN (17.5%) zygotes, while 1PN zygotes showed a significantly lower rate (7.0%) compared to both 0PN and 2PN. Clinical pregnancy, ectopic pregnancy, implantation, and live birth rates were similar among single blastocyst frozen embryo transfers (FET) of 0PN, 1PN, and 2PN. Additionally, no significant differences were observed between single- and double-blastocyst FET of 0PN and 2PN. Our findings suggest that 0PN and 2PN zygotes have comparable developmental potential, while 1PN embryos exhibit lower developmental potential. Blastocyst FET outcomes appear similar among 0PN, 1PN, and 2PN zygotes.

Effects of total gonadotropin dose on embryo quality and clinical outcomes with AMH stratification in IVF cycles: a retrospective analysis of 12,588 patients

BackgroundRecent studies about the effect of gonadotropin (Gn) dose on the clinical outcomes of IVF are still controversial, and no studies have analyzed the relationship between Gn dose and embryo quality. Since AMH is a strong predictor of oocyte quality, we aim to evaluate the relationship between total Gn dose and embryo quality and clinical outcomes at different AMH levels in IVF cycles.MethodsA total of 12,588 patients were enrolled in the retrospective study. The included cycles were categorized by serum AMH levels (AMH ≤ 1 ng/ml, 1 ng/ml < AMH ≤ 3 ng/ml, 3 ng/ml < AMH ≤ 5 ng/ml, AMH > 5 ng/ml), total Gn dosage (< 1875 IU, 1875–3750 IU and ≥ 3750 IU) and female age (< 35 years and 35–42 years). The embryo quality and clinical outcomes were the measure outcomes.ResultsThe top-day3 embryos rate decreased with the increase of total Gn dose in nearly all age and AMH subgroups, but this trend was not obvious in the AMH > 5 ng/ml group and AMH ≤ 1 ng/ml group. The blastocyst formation rate and high-quality blastulation rate had a negative relationship with Gn does for women aged < 35 years in the AMH ≤ 5 ng/ml groups, except for the AMH > 5 ng/ml group (P < 0.001). However, when women were 35–42 years old, regardless of AMH levels, the blastocyst formation rate and high-quality blastulation rate decreased as Gn dose increased. Clinical outcomes (implantation rate, clinical pregnancy rate and live birth rate) decreased with the increase of Gn dose in all ages and AMH stratifications.ConclusionsThe total dose of Gn may have different effects on embryo quality at different serum AMH levels, and the negative effects of total dose of Gn on clinical outcomes may be realized by impairing both embryo quality and endometrium.

The impact of clinical and laboratory parameters on clinical pregnancy and live birth rates in fresh cycles: a retrospective study of 9608 high-quality cleavage-stage embryos

BackgroundEvidence from the Istanbul consensus workshop suggests correlations between morphological parameters and embryo developments. 8-cell embryos are the best blastomere stage on day 3. No good quality evidence exists to support high-quality embryonic selection following blastulation and clinical outcomes. This study aimed to investigate the factors that affect blastocyst formation, blastocyst quality, and clinical outcomes of high-quality cleavage-stage embryos in fresh cycles.MethodsThis study was a retrospective analysis of 9608 high-quality cleavage-stage embryos from 2987 couples between January 2017 to June 2021, namely 1520 embryos categorized as “812” (8-cell, grade 2, mild fragmentation), 2961 as “821” (8-cell, grade 2, mild asymmetry), 896 as “711” (7-cell, grade 1), and 517 as “911” (9-cell, grade 1) compared with 3714 embryos categorized as “811” (8-cell, grade 1). The primary outcomes were clinical pregnancy rate (CPR) and live birth rate (LBR). Blastulation rate (BR), available late blastocyst rate (ABR) and high-quality late blastocyst rate (HBR) were secondary outcome measures.ResultsBR, ABR, and HBR had significant differences among the five groups (P < 0.001), while CPR and LBR were also significantly different in cleavage-stage fresh transfer (P < 0.01). The multivariable multilevel logistic regression analysis revealed a significant association between cell number, cell size, blastocyst development and clinical outcomes. For 7 to 9-cell highest-quality embryo, mild fragmentation and more blastomeres were more conducive to blastocyst formation and clinical outcomes. While cleavage-stage embryos developed into blastocysts, the negative impact of their initial morphology on clinical outcomes would be erased.ConclusionsOur study firstly evaluated blastocyst development and clinical outcomes of high-quality cleavage-stage embryos in fresh cycles, with rankings of 811, 812, 911, 821, and 711. We found the initial morphological characteristics of the high-quality cleavage-stage embryos did not adversely impact clinical outcomes, even as they progressed to the blastocyst stage.

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.

The ovarian stimulation regimen does not affect aneuploidy or blastocyst rate

Could the total dose (<3000 IU or ≥3000 IU) and type of exogenous gonadotrophin (i.e. recombinant FSH and/or human menopausal gonadotrophin [HMG]) influence aneuploidy and blastulation rates and produce different reproductive outcomes? This retrospective, observational, multicentre cohort study included a total of 8466 patients undergoing IVF using autologous oocytes and preimplantation genetic testing for aneuploidies. Participants were divided according to the dosage of total gonadotrophins and stratified by maternal age. The aneuploidy rates, pregnancy outcomes and cumulative live birth rates (CLBR) were similar among women who received total gonadotrophin dosages of <3000 or ≥3000 IU. No statistical differences were reported in the blastulation rate with lower or higher gonadotrophin dosages. Women receiving a higher amount of HMG during ovarian stimulation had a lower aneuploidy rate (P = 0.02); when stratified according to age, younger women with a higher HMG dosage had lower aneuploidy rates (P< 0.001), while no statistical differences were observed in older women with higher or lower HMG dosages. No significant differences were observed in IVF outcomes or CLBR. High doses of gonadotrophins were not associated with rate of aneuploidy. However, an increased fraction of HMG in younger women was associated with a lower aneuploidy rate. The study demonstrated that the total gonadotrophin dosage did not influence aneuploidy, reproductive outcomes or CLBR. The increased gonadotrophin and HMG dosages used for ovarian stimulation did not precede aneuploidy, and the use of HMG should be evaluated on a case-by-case basis, according to the individual's characteristics and infertility type.

Unhealthy air quality secondary to wildfires is associated with lower blastocyst yield

To study the impact of unhealthy air quality from the 2020 Oregon wildfires on outcomes for patients undergoing invitro fertilization (IVF) treatment. A retrospective cohort study. A university-based fertility clinic. Subjects were undergoing IVF treatment from the 6 weeks preceding the wildfires through a 10-day exposure period. Cohorts were classified on the basis of whether subjects experienced patient and/or laboratory exposure to unhealthy air quality. Patient exposure was defined as at least 4 days of ovarian stimulation overlapping with the exposure, and laboratory exposure was defined as at least 2 days of IVF treatment and embryogenesis overlapping with the exposure. The unexposed cohort consisted of remaining subjects without defined exposure, with cycles in the 6 weeks preceding the wildfires. As some subjects had dual exposure and appeared in both patient and laboratory exposure cohorts, each cohort was separately compared with the unexposed control cohort. A 10-day period of unhealthy air quality caused by smoke plumes from a wildfire event. The primary outcome was the blastulation rate. Secondary outcomes included fertilization rate, number of blastocysts obtained, and cycles with no blastocysts frozen or transferred. Sixty-nine subjects underwent ovarian stimulation and IVF treatment during the 6 weeks preceding the wildfires through the 10-day period of unhealthy air quality. Of these, 15 patients were in the laboratory exposure cohort, 16 were in the patient exposure cohort, and 44 were unexposed. Six subjects appeared in both laboratory and patient exposure cohorts. Although neither exposure cohort had significantly decreased blastulation rate compared with the unexposed, the median number of blastocysts obtained was significantly lower in the laboratory exposure cohort than the unexposed group (2 [range 0-14] vs. 4.5 [range 0-21], respectively). The laboratory exposure cohort had significantly more cycles with no blastocysts obtained (3/15 [20%] vs. 1/44 [2%]). There were no significant differences in IVF treatment outcomes between patient exposure and unexposed cohorts. These findings persisted after controlling for age. There were no significant differences in pregnancy outcomes observed after embryo transfer between the exposure group and the unexposed group. For a cohort of patients undergoing IVF treatment, an acute episode of outside wildfire smoke exposure during fertilization and embryogenesis was associated with decreased blastocyst yield.

Lipopolysaccharide-binding protein in follicular fluid is associated with the follicular inflammatory status and granulosa cell steroidogenesis in dairy cows.

Metabolic stress and subsequent hepatic dysfunction in high-producing dairy cows are associated with inflammatory diseases and declining fertility. Lipopolysaccharide (LPS)-binding protein (LBP) is produced by hepatocytes and controls the immune response, suggesting that it is involved in the pathophysiology of inflammation-related attenuation of reproductive functions during metabolic stress. This study investigated the effect of LBP on the inflammatory status, oocyte quality, and steroidogenesis in the follicular microenvironment of dairy cows. Using bovine ovaries obtained from a slaughterhouse, follicular fluid and granulosa cells were collected from large follicles to evaluate the follicular status of metabolism, inflammation, and steroidogenesis. Cumulus-oocyte complexes were aspirated from small follicles and subjected to in vitro embryo production. The results showed that follicular fluid LBP concentrations were significantly higher in cows with fatty livers and hepatitis than in those with healthy livers. Follicular fluid LBP and LPS concentrations were negatively correlated, whereas LPS concentration showed a positive correlation with the concentrations of non-esterified fatty acids (NEFA) and β-hydroxybutyric acid in follicular fluid. The blastulation rate of oocytes after in vitro fertilization was impaired in cows in which coexisting large follicles had high NEFA levels. Follicular fluid NEFA concentration was negatively correlated with granulosa cell expression of the estradiol (E2) synthesis-related gene (CYP19A1). Follicular fluid LBP concentration was positively correlated with follicular fluid E2 concentration and granulosa cell CYP19A1 expression. In conclusion, follicular fluid LBP may be associated with favorable conditions in the follicular microenvironment, including low LPS levels and high E2 production by granulosa cells.

ICSI using testicular spermatozoa after failure of ICSI with ejaculated spermatozoa could be a good choice: A propensity score-matched cohort study.

Ejaculated spermatozoa are considered to possess a higher fertilisation potential than testicular spermatozoa. In selected cases, the use of testicular spermatozoa from non-azoospermic infertile men resulted in a higher implantation and pregnancy rate than the use of ejaculated spermatozoa. The primary objective was to compare the live birth rate and cumulative live birth rate between couples with failed intracytoplasmic sperm injection procedure using ejaculated spermatozoa who subsequently had an intracytoplasmic sperm injection cycle with testicular spermatozoa and those who subsequently had an intracytoplasmic sperm injection cycle with ejaculated spermatozoa. The secondary objective was to determine the indications for the use of testicular spermatozoa after intracytoplasmic sperm injection failure with ejaculated spermatozoa. A retrospective study of matched couples using propensity score matching analysis was performed. After an intracytoplasmic sperm injection failure (cycle_1), intracytoplasmic sperm injection with either ejaculated spermatozoa (ejaculated sperm group), or testicular spermatozoa (testicular sperm group), was performed (cycle_2). The matching was on intracytoplasmic sperm injection performed in cycle_1 according to spermatozoa used (testicular or ejaculated) in cycle_2. Logistic regression was used to evaluate the influence of sperm origin on cumulative live birth rate. Univariate analysis on parameters of cycle_1 was used to identify the prognostic factors to propose an intracytoplasmic sperm injection with testicular spermatozoa in case of cycle_1 failure. The study outcomes were live birth rate and cumulative live birth rate. Among the 6034 couples available, 63 were selected to constitute the testicular sperm group and 63 were selected by propensity score matching to constitute the ejaculated sperm group. After matching, the DNA fragmentation index was higher in the testicular sperm group (13.43%±9.65% vs. 8.93%±4.47%, p=0.013); no significant difference was observed for the fertilisation rate, the number of obtained embryos, blastulation rate and frozen embryo rate. In cycle_2, the live birth rate was higher in the testicular group (22.2% vs. 0.0%, p<0.001), as was the cumulative live birth rate (25.4% vs. 6.3%, p=0.065). The prognostic factors identified for the proposal of intracytoplasmic sperm injection procedure with testicular spermatozoa after intracytoplasmic sperm injection failure with ejaculated spermatozoa were: teratozoospermia, cryptozoospermia and high DNA fragmentation index. According to the present study and current knowledge, the use of testicular spermatozoa after failed intracytoplasmic sperm injection procedure in non-azoospermic men could be proposed instead of sperm donation in case of high sperm DNA fragmentation index, cryptozoospermia and teratozoospermia. A good oocyte response to ovarian stimulation during the previous assisted reproductive technology attempt will increase the chance of success. Although the main limitation of the current study is its retrospective nature, the use of the propensity score matching to perform causal inference study increases its reliability. The present study supports that the use of testicular spermatozoa outside the classical indication of azoospermia is a good option when the indication is well established. However, before proposing a testicular biopsy, an improvement in sperm characteristics should be considered by treating the causes of sperm alteration.

#88 : Correlation Between AMH Levels and Aneuploidy Rates in Patients Over 40 Years of Age: A Novel Finding

Background and Aims: AMH is a crucial biomarker of ovarian reserve and predictor of ovarian response to stimulation. Ovarian reserve is inversely proportional to women’s age, consequently lower AMH levels are seen in older women particularly at [Formula: see text]40 yrs of age. A correlation between age and embryo aneuploidy was established but not with AMH level. We hypothesize that older women with good ovarian reserve produce less aneuploid embryos and produce the same blastocyst rate. Method: A retrospective study was undertaken involving 507 IVF cycles in patients aged [Formula: see text]40 years who had ICSI with PGT-A using NGS. The cycles were categorized into three main cohorts based on the patient’s AMH level: low (&lt; 1.1 ng/mL), moderate (&gt;1.1 to [Formula: see text] 3.0 ng/mL) and high (&gt; 3.0 ng/mL). Each cohort was subdivided into 3 age sub-cohorts as follow: (40 to &lt;42), (42 to &lt;44), and (&gt;44). Our primary outcome was aneuploidy rate whereas the secondary outcome was blastulation rate. Results: The results will be mentioned in sequence: low AMH, moderate AMH and, high AMH levels. In the [Formula: see text]40 to &lt; 42 age group, the aneuploidy rates were 78.2%, 72.3% and, 67.2%, respectively (n = 232; p = 0.027). The blastulation rates were 53.8%, 47.7% and, 48.5% respectively (p&gt; 0.05). In the [Formula: see text]42 and &lt;44 age group, the aneuploidy rates were 89.2%, 83.2% and, 81.5% respectively (n = 136 p=0.039). The blastulation rates were 45.2%, 46.2% and, 43.8% respectively p&gt; 0.05. In the [Formula: see text]44 age group, the aneuploidy rates were 93.9%, 91.2% and, 89.4% respectively (n = 139; p = 0.043). The blastulation rates were 41.2%, 39.2% and, 38.8% respectively (p&gt;0.05). Conclusion: The aneuploidy rates rises when AMH levels decreases; whereas the blastulation rates was not associated with AMH levels in the population studied.

#154 : The Creation of a New Embryo Culture Medium with 31 Optimal Components Derived from Human Oviduct Fluid: A Prospective Multicenter Randomised Trial

Background and Aims: Although culture conditions were recognized as key points, an optimal culture medium formulation, which closely resembles natural human oviduct fluid (HOF), has not been obtained yet. Our aim is to make an ideal embryo culture medium close to HOF components, and to evaluate the quality of this medium with embryo quality and clinical outcomes in assisted reproductive technology (ART) by a prospective randomized controlled trial (RCT). Method: Study I: HOF was collected laparoscopically from patients (n = 28) with normal pelvic findings. According to HOF analysis results, the new medium ‘OVIT’ was designed. Study II: Embryos (2 pronuclei (2PN) = 9,633) were assigned from 1,435 patients. The blastulation rate (BR), good BR (gBR), utilized (transferred/cryo-preserved) BR (uBR), pregnancy rate (PR), and miscarriage rate (MR) were compared between the OVIT and control groups by RCT. Study III: The neonatal healthiness who were concepted by ART using OVIT and /or control medium was compared between two groups. Results: The novel medium ‘OVIT’ was produced according to 31 HOF components. The concentrations of essential amino acids (e-AAs) were lower in OVIT than in current media, yet the opposite was true for non-e-AA concentrations. gBR and uBR were higher in the OVIT group than in the control group. In the older female group, gBT and uBR were significantly higher in the OVIT group. The neonatal conditions of those who were born by ART using OVIT and/or control medium were similar. Conclusion: HOF components and AA concentrations were analysed, and the results indicated numerous differences between OVIT and current media. This medium provides a better culture condition and improves embryo quality. This study has found novel perspectives for older patients having difficulties in producing embryos of good quality and indicated the superior quality of OVIT compared with the quality of traditional media.

#27 : A Sibling Study: How Individual Culture Strategy Affects Embryo Ploidy Status in IVF/PGT-A Cycles?

Background and Aims: The result of individual and group culture embryo studies remained conflicting. Several studies assumed that the blastulation rate was lower when culturing embryos separately. Meanwhile, there has been little information on whether individual culture can affect embryo ploidy status. This study was designed to find a relationship between culture strategies and the embryo’s ploidy status in IVF/PGT-A cycles. Method: 91 IVF/PGT-A cycles with at least four zygotes enrolled in a randomized control trial from September 2021 to June 2022 in Tam Anh general hospital. Zygotes from each cycle were randomly assigned to individual culture(10 ul/drop) or group cultured of 3-5 zygotes (50 ul/drop). Among those, 280 blastocysts derived from biopsy-all cycles (137 from individual culture and 143 from group culture) and already determined for ploidy status were to be analyzed. Information of the embryo, including maternal age, paternal age, day of the biopsy, embryo quality (excellent, good, fair, poor), blastocoele expansion (2-6), ICM score (A, B, C), TE score (A, B, C), and PGT-A result (euploid, aneuploidy or else) was collected and analyzed along with the type of culture. Simple and multiple regression analyses using generalized linear mixed model were applied to evaluate the relation between embryo ploidy status as the dependent variable and independent factors. Results: The rates of euploid, aneuploidy and other results of embryos cultured individually were 39.4%, 37.2%, and 23.4%, respectively. These results did not yield a significant difference compared with the cultured group. There was no relationship between culture strategies and the aneuploidy rate of the embryos (OR = 1.16, CI 0.64-2.13). However, biopsy days 6-7 and decreasing grade of TE might be related factors for a higher rate of aneuploidy. Conclusion: Culture embryos individually or in groups hadnot yet shown any effect on embryo’s ploidy status.

Embryological characteristics and clinical outcomes of oocytes with different degrees of abnormal zona pellucida during assisted reproductive treatment.

Abnormalities in the zona pellucida (ZP) adversely affect oocyte maturation, embryo development and pregnancy outcomes. However, the assessment of severity is challenging. To evaluate the effects of different degrees of ZP abnormalities on embryo development and clinical outcomes, in total, 590 retrieval cycles were scored and divided into four categories (control, mild, moderate and severe) based on three parameters: perivitelline space, percentage of immature oocytes and percentage of oocytes with abnormal morphology. As the severity of abnormal ZP increased, both the number of retrieved oocytes and mature oocytes decreased. The fertilization rate did not differ significantly among groups. The rates of embryo cleavage and day-3 high-quality embryos in the mild group and the moderate group did not vary significantly between the two groups but were significantly higher than those in the severe group. The blastulation rates of the abnormal ZP groups were similar; however, they were lower than those of the control group. Moreover, the cycle cancellation rate of the severe abnormal ZP group was as high as 66.20%, which was significantly higher than that of the other three groups. Although the rates of cumulative clinical pregnancy and live births were lower than those in the control group, they were comparable among the abnormal ZP groups. There were no differences in the neonatal outcomes of the different groups. Together, ZP abnormalities show various degrees of severity, and in all patients regardless of the degree of ZP abnormalities who achieve available embryos, there will be an opportunity to eventually give birth.

Intra-patient analysis of individual weight gain or loss between IVF cycles: cycle now and transfer later.

What is the impact of clinically significant weight change on outcomes related to IVF cycle performance? While individual weight loss did not significantly impact ovarian response to stimulation or other cycle outcome parameters in our study, some positive associations were found for individual weight gain. The role of weight-change in patients undergoing IVF has been largely studied by comparing weight loss in different cohorts of patients stratified by a static BMI. Specifically, obesity has been extensively studied in relation to its negative effects on assisted or unassisted conception outcomes and ovulatory function. Previous research has shown conflicting results, while BMI, which is commonly used as a marker of obesity, may not accurately reflect the underlying factors affecting fertility in obese patients. This study utilized a retrospective within-patient repeated measurement analysis design to assess the impact of weight change on IVF outcomes in cycles where all embryos were cryopreserved at the blastocyst stage for transfer at a later date. The study was conducted at an academically affiliated fertility center. The data included 961 women who underwent at least two IVF cycles between December 2014 and June 2020, with documented short-term weight gain (n = 607) or weight loss (n = 354) within 1year from their initial IVF cycle. Multivariable generalized estimating equations (GEE) and generalized linear mixed models (GLMM) were employed to assess associations between weight change and outcomes across cycles. The multivariable models indicated that weight loss did not show any significant associations with the numbers of oocytes retrieved, or mature oocytes, the fertilization rate or the blastulation rate. However, weight gain demonstrated a minor positive association with the number of oocytes retrieved in both GEE models (coefficient: 0.01, 95% CI: 0.00-0.01) and GLMM models (0.01, 95% CI: 0.01-0.00). There was also a potential increase in the fertilization rate with weight gain, as indicated by a positive coefficient in both GEE models (coefficient: 0.01, 95% CI: 0.00-0.02) and GLMM models (coefficient: 0.01, 95% CI: 0.00-0.01). However, the association between weight gain and the embryo blastulation rate was not statistically significant in any model. This study focused on cycle performance parameters instead of reproductive outcomes, which restricted our ability to evaluate the impact of weight change on cumulative live birth rates. Additionally, the study did not account for variables such as stimulation protocols, potentially introducing confounding factors and limiting the generalizability of the results. Although obesity is associated with adverse obstetrical risks, there is less evidence of adverse reproductive outcomes in IVF cycles. We therefore recommend that an IVF cycle should not be delayed due to weight, so that the patient is not adversely affected by increasing age. The IVF cycle should aim to freeze all embryos, so that embryo transfer can then occur after weight loss, so as to limit the recognized obstetrical risks. The study was not funded and there were no competing interests. N/A.

Enhancement of preimplantation mouse embryo development with optimized in vitro culture dish via stabilization of medium osmolarity.

We evaluated the efficacy of the newly developed optimized in vitro culture (OIVC) dish for cultivating preimplantation mouse embryos. This dish minimizes the need for mineral oil and incorporates microwells, providing a stable culture environment and enabling independent monitoring of individual embryos. Mouse pronuclear (PN) zygotes and two-cell-stage embryos were collected at 18 and 46 hours after human chorionic gonadotropin injection, respectively. These were cultured for 120 hours using potassium simplex optimized medium (KSOM) to reach the blastocyst stage. The embryos were randomly allocated into three groups, each cultured in one of three dishes: a 60-mm culture dish, a microdrop dish, and an OIVC dish that we developed. The OIVC dish effectively maintained the osmolarity of the KSOM culture medium over a 5-day period using only 2 mL of mineral oil. This contrasts with the significant osmolarity increase observed in the 60-mm culture dish. Additionally, the OIVC dish exhibited higher blastulation rates from two-cell embryos (100%) relative to the other dish types. Moreover, blastocysts derived from both PN zygotes and two-cell embryos in the OIVC dish group demonstrated significantly elevated mean cell numbers. Use of the OIVC dish markedly increased the number of cells in blastocysts derived from the in vitro culture of preimplantation mouse embryos. The capacity of this dish to maintain medium osmolarity with minimal mineral oil usage represents a breakthrough that may advance embryo culture techniques for various mammals, including human in vitro fertilization and embryo transfer programs.

Seeking arrangements: cell contact as a cleavage-stage biomarker

Research questionWhat can three-dimensional cell contact networks tell us about the developmental potential of cleavage-stage human embryos? DesignThis pilot study was a retrospective analysis of two Embryoscope imaging datasets from two clinics. An artificial intelligence system was used to reconstruct the three-dimensional structure of embryos from 11-plane focal stacks. Networks of cell contacts were extracted from the resulting embryo three-dimensional models and each embryo's mean contacts per cell was computed. Unpaired t-tests and receiver operating characteristic curve analysis were used to statistically analyse mean cell contact outcomes. Cell contact networks from different embryos were compared with identical embryos with similar cell arrangements. ResultsAt t4, a higher mean number of contacts per cell was associated with greater rates of blastulation and blastocyst quality. No associations were found with biochemical pregnancy, live birth, miscarriage or ploidy. At t8, a higher mean number of contacts was associated with increased blastocyst quality, biochemical pregnancy and live birth. No associations were found with miscarriage or aneuploidy. Mean contacts at t4 weakly correlated with those at t8. Four-cell embryos fell into nine distinct cell arrangements; the five most common accounted for 97% of embryos. Eight-cell embryos, however, displayed a greater degree of variation with 59 distinct cell arrangements. ConclusionsEvidence is provided for the clinical relevance of cleavage-stage cell arrangement in the human preimplantation embryo beyond the four-cell stage, which may improve selection techniques for day-3 transfers. This pilot study provides a strong case for further investigation into spatial biomarkers and three-dimensional morphokinetics.

Is there any difference on fertilization and blastulation rates of sibling oocyte activated by piezo-electric or ca ionophore application?

Is there any difference on fertilization and blastulation rates of sibling oocyte activated by piezo-electric or ca ionophore application?

The impact of increasing body mass index on in vitro fertilization treatment, obstetrical, and neonatal outcomes

BackgroundCiting the risks of administering anesthesia to patients with obesity, few fertility centers offer IVF as a treatment modality for patients with BMIs ≥ 40 kg/m2. Although previous studies have assessed clinical pregnancy and cumulative live birth rates in patients who spontaneously conceive with BMIs ≥ 50 kg/m2, there is a paucity of IVF, obstetric, and neonatal outcome data in patients with severe obesity who conceive after IVF. ObjectiveTo evaluate the impact of increasing BMI on IVF, obstetrical, and neonatal outcomes in patients with obesity undergoing IVF. Study DesignThis was a retrospective cohort study within an academic fertility center including 2,069 fresh IVF/ICSI and frozen embryo transfer cycles from January 1, 2012 to April 30, 2020: this cohort was utilized to determine IVF treatment outcomes. A second embedded cohort of 867 fresh IVF/ICSI and frozen embryo transfer cycles which resulted in ongoing clinical pregnancies and deliveries within a single tertiary hospital system was utilized to determine pregnancy, maternal, and neonatal outcomes. All patients with a BMI ≥ 40 kg/m2 underwent consultation with a maternal fetal medicine specialist prior to starting treatment and a preoperative evaluation with an anesthesiologist prior to oocyte retrieval. Cycles were grouped by body mass index at cycle start: BMI 30-34.9 kg/m2, BMI 35-39.9 kg/m2, BMI 40-44.9 kg/m2, BMI 45-49.9 kg/m2, and BMI > 50 kg/m2. Log-binomial Poisson regression and Poisson regression with an offset models were performed with BMI 30-34.9 kg/m2 as the reference group, adjusting for potential confounders including oocyte age, patient age, embryo quality, transfer type, and coexisting comorbidities. The primary outcome was live birth rate. Secondary outcomes included fertilization rate, blastulation rate, miscarriage rate, incidence of preeclampsia with severe features, gestational diabetes, labor induction, cesarean section, preterm delivery, and birth weight. ResultsThere were 2,069 fresh IVF/ICSI and frozen embryo transfer cycle starts from January 1, 2012 to April 30, 2020. Of these, 1008 cycles were in the BMI 30-34.9 kg/m2 group, 547 in the BMI 35-39.9 kg/m2 group, 277 in the BMI 40-44.9 kg/m2 group, 161 in the BMI 45-49.9 kg/m2 group, and 76 in the BMI > 50 kg/m2 group. Live birth rate was not significantly different between groups. The BMI > 50 kg/m2 group was significantly more likely to experience preeclampsia with severe features when compared to the BMI 30-34.9 kg/m2 group (aRR 2.75, 95% CI 1.13-6.67). Fertilization rate, blastulation rate, miscarriage rate, incidence of gestational diabetes, labor induction, cesarean section, preterm delivery, and neonatal birth weights were not significantly different between groups. Conclusion(s): Among patients with BMIs 30-60 kg/m2 who conceived via IVF and received comprehensive prenatal care at a tertiary care hospital, IVF, obstetrical, and neonatal outcomes were largely comparable. These data support a collaborative care approach with maternal fetal medicine specialists and skilled anesthesiologists, reinforcing the notion that IVF should not be withheld as a treatment modality from patients with obesity.

Impact of baseline luteinizing hormone (LH) level and anti-Müllerian hormone (AMH) on pregnancy outcomes in polycystic ovary syndrome (PCOS) patients undergoing IVF/ICSI cycle in antagonist protocol

Introduction: Polycystic ovary syndrome (PCOS) is a complex endocrine and metabolic disorder often associated with infertility. There are several factors that could impair the pregnancy outcomes of PCOS patients. The latter endocrine profile along with high baseline luteinizing hormone (LH), high LH/follicle-stimulating hormone ratio, as well as high anti-Müllerian hormone (AMH) levels appear detrimental for patients undergoing In vitro fertilization (IVF)/intracytoplasmic sperm injection (ICSI). The objective of this study is to evaluate the impact of baseline LH and AMH levels on pregnancy outcomes in PCOS patients performing IVF/ICSI cycles in an antagonist protocol. Materials and methods: The current investigation is a retrospective cohort study conducted at IVF Tiziri Center in Algiers on PCOS patients undergoing IVF/ICSI from January 2017 to March 2021. According to the Rotterdam criteria, patients aged between 23 and 42 years were divided into 2 groups: group 1 (G1) =91 patients; group 2 (G2) =100 all patients had an LH level &gt;10 IU/L. Both groups had AMH levels &gt;5 ng/mL. Controlled ovarian stimulation was done with a fixed antagonist protocol, as for the ovulation it was triggered with human chorionic gonadotropin or gonadotropin-releasing hormone agonist. Fresh and frozen embryo transfers were, thereafter, evaluated. Concerning the statistical analysis, it was done with SPSS software. Results: The mean LH level was 15.16±7.08 for G1 versus 5.7±1.86 for G2 (P=10−6), mean LH/follicle-stimulating hormone ratio 2.71±1.36 in G1 versus 1.05±0.44 (P=10−6) and mean AMH level was 9.27±3.90 for G1 versus 7.53±2.76 for G2 with (P&lt;10−3). The mean follicles over 11 mm in diameter, estradiol (E2) level, and progesterone level on the trigger day did not differ significantly between G1 and G2. Also, the mean of oocytes and metaphase II oocytes retrieved fertilization, and blastulation rate were similar for both groups. The pregnancy rate for G1 was 40.6% versus 54% (P&gt;0.05); the ongoing pregnancy rate (OPR) was 28.6% in G1 versus 47% in G2 (P=0.078); and the miscarriage rate was 32.4% for G1 versus 12.9 for G2 (P=0.072). A univariate analysis was performed with all confounder factors supposed to impact pregnancy outcomes: AMH level did not impact pregnancy outcomes, whereas LH level &lt;11 IU/L has been determined as a threshold impacting positively on OPR with odds ratio=2.01 (95% CI: 1.05–3.87). After a multivariate logistic regression including con-founders, 3 factors significantly influence OPR: LH, E2, and infertility with known etiology. Conclusion: The high baseline LH level impairs pregnancy outcomes in PCOs patients undergoing an antagonist protocol IVF/ICSI cycle. The identification of PCOS patients by their endocrine profile enables the establishment of a prognosis for council patients and a plan for an individualized therapeutic strategy.