P–197 Two different strategies for embryo culture and selection: time-lapse with single-step medium and conventional incubator with sequential media. Are there differences in clinical results?

2021 ◽  
Vol 36 (Supplement_1) ◽  
Author(s):  
C Alber. Rodriguez ◽  
M Valera ◽  
L Bori ◽  
F Meseguer ◽  
L Alegre ◽  
...  

Abstract Study question Is there a significant difference in the clinical results of embryos cultured in time-lapse systems with single-step medium and conventional benchtop incubators with sequential media? Summary answer Embryos cultured in time-lapse systems and single-step media are more likely to achieve an ongoing pregnancy and have higher implantation rates than those cultured otherwise. What is known already One of the strategies for embryo culture in IVF consisted in conventional benchtop incubators combined with sequential culture media (CI-Seq). New generation time-lapse systems provide useful information on the morphokinetics of embryo development, but also a stable culture environment where embryos can develop undisturbed until blastocyst stage when paired with single-step culture media (TLS-SS). These features have the potential to improve embryo development and selection. Nonetheless, there is inconclusive evidence of whether this new culture strategy has a significant effect on clinical results of ICSI treatments. Studies on the matter are heterogeneous and reduced in both number and sample size. Study design, size, duration Unicentric retrospective cohort study. We compared the results of 11471 blastocyst transferences from 10276 ICSI treatments performed during 4 consecutive years, where embryos were cultured either on CI with sequential media (N = 5255) or a TLS with single-step medium (N = 5021). 3922 of the totals were fresh embryo transfers (ET) and 7549 frozen-thawed ET. We compared the implantation rate (IR) and ongoing pregnancy rate (OGPR) in both study groups, stratifying by ovum origin. Participants/materials, setting, methods Three models of TLS were used for embryo culture: EmbryoScope, EmbryoScope Plus (Vitrolife) and GERI (Genea Biomedx), as well as one CI (ASTEC). Sequential media: Cook, Origio, Vitrolife; Single-step media: Gems, Irvine, Life Global. Embryo scoring and selection was performed by ASEBIR criteria in the CI group, and by morphological and morphokinetic assessment for embryos cultured in TLS. Embryos were extracted from the CI only for media change. Statistical analysis: ANOVA tests and Logistic regressions. Main results and the role of chance A general Logistic Regression was performed, including egg origin, PGT-A and culture strategy to explain their impact in OGPR. Egg origin (OR = 1,094 (95%CI: 1,015–1,179); P = 0,019) and culture strategy (OR = 1,141 (95%CI: 1,060–1,229); P < 0,001) were statistically significant, which confirms the need for stratification due to the heterogeneity of the groups. The total IR in the TLS-SS group was 54,68±48,84%, significantly higher than that of CI-Seq (49,18±47,91%; P < 0,001). In ovum-donation treatments, a complete Logistic Regression for OGPR, with all typical confounding variables (age, BMI, nº oocytes, fresh/frozen transfer, number and day of ET) resulted in an OR = 1,187 (95%CI: 1,074–1,313; P = 0,001) favoring culture in TL-SS. IR in these treatments were 61,98±47,68% in TL-SS vs 55,08±46,58% in CI-Seq (P < 0,001) in fresh transfers and 51,48±48,91% in TL-SS vs 44,39±47,67% in CI-Seq (P < 0,001) in frozen-thawed ET. In autologous treatments with PGT a similar regression yielded an OR = 1,055 (95%CI: 0,889–1,252; P = 0,542) for culture strategy. The IR of genetically tested ET was not significantly different: 53,08±49,49% for TL-SS, 50,90±49,07% for CI-Seq, P = 0,246. In autologous procedures without PGT, culture strategy was not significant for OGPR (OR = 0,998 (95%CI: 0,835–1,191), P = 0,979) nor IR of fresh (49,75±48,91% TL-SS vs 44,23±47,36% CI-Seq; P = 0,081) nor frozen-thawed transferences (50,77±48,33% TL-SS vs 50,67±47,33% CI-Seq; P = 0,970). Limitations, reasons for caution After fertilization check, embryos were evaluated exclusively on D5/6. On D3, embryos cultured in CI were taken out only for a quick media change, but not for evaluation, and all handling was done in isolette cabins with controlled environmental conditions. Being a retrospective study, there is high variability in population. Wider implications of the findings: A more homogenous prospective study, including comparison in life-birth rates, is necessary to extract final conclusions. However, our results suggest that the introduction of TLS and SS media in IVF laboratories might be a valid strategy to increase clinical results, especially in fresh embryo, thanks to an improved embryo selection. Trial registration number Not applicable

2021 ◽  
Vol 36 (Supplement_1) ◽  
Author(s):  
M.D.L.Á. Valera Cerdá ◽  
C Albert ◽  
L Bori ◽  
J Marcos ◽  
Z Larreategui ◽  
...  

Abstract Study question Does culture in high relative humidity conditions (HC) improve pregnancy rates when using a time-lapse system (TLS) and single-step (SS) culture medium? Summary answer Using an integrated-TLS and SS medium, culture under HC increases the likelihood of embryos to achieve a pregnancy with respect to those cultured in DC. What is known already Many variables affect embryo development, and need to be precisely tuned in every IVF laboratory, especially inside the incubators. TLS provide stability during embryo culture, which is a well-known key factor for a proper embryo development. The humidity content of culture atmosphere is especially relevant in order to avoid oscillations in culture media osmolality. It has been previously reported that culture under HC has a significant effect on embryo quality and morphokinetics. However, studies assessing the effect of HC in clinical outcome are rare and inconclusive, mostly due to the variability in the incubator device used and insufficient sample size. Study design, size, duration The present is a retrospective study performed over 1624 ICSI treatments from 3 fertility clinics from December 2017 to October 2020. Zygote cohorts were randomly assigned to dry (N = 794) or humid conditions (N = 830). It includes autologous treatments with (N = 555) and without (N = 368) pre-implantation genetic testing (PGT) and egg donation treatments (N = 701). Following selection by combining morphological and morphokinetic criteria, 1611 mostly single embryo transfers (92%) were performed, 779 from DC and 832 from HC. Participants/materials, setting, methods Stimulation, oocyte pickup and fertilization were performed according to the standard procedures of the clinic. We used a GERI incubator (Genea Biomedx), with 6 separated chambers for individual patients, 3 of them configured to work in DC, and 3 in HC. Embryos were cultured in specific 16-well GERI trays with single-step Gems® culture medium (Genea Biomedx). The effect of HC in pregnancy rate was assessed by multivariate logistic regression and Pearson Chi Square Test. Main results and the role of chance Types of treatment and patient demographics were homogeneously distributed in the two study groups. Mean patient age was 39.88±4.47 years, BMI: 23.54±4.21 Kg/m2 and number of correctly fertilized oocytes: 7.86±3.87. A logistic regression was performed, including other possible affecting factors: ovum age and origin, transfer day, fresh or frozen-warmed embryo transfer, number of transferred embryos and the use of PGT. Said analysis revealed that embryos cultured in HC are more likely to achieve a pregnancy than those cultured in DC (OR = 1.30, 95% CI (1.05-1.59), p=0.014). Pregnancy rate was significantly higher in HC (66.7%) than in DC (60.9%) in the total embryo transfers (p = 0.017). Pregnancy rate was also higher in HC in fresh embryo transfers (68.6% in HC vs 63.2% in DC; p = 0.133) and frozen-thawed transfers (65.2% in HC vs 59.1% in DC; p = 0.062), although differences were not statistically significant due to the reduced sample size. Stratifying the results, the significant difference remained in transfers belonging to autologous cycles (68.4% HC vs 56.5% in DC; p = 0.030) and in treatments in which PGT was performed (67.1% HC vs 56.0% in DC; p = 0.023), but the difference in egg donation procedures was not statistically significant (66.4% in HC vs 64.7% in DC, p = 0.577). Limitations, reasons for caution This is a retrospective analysis performed over the clinics’ treatments, so it might be compromised by some bias, although multivariable analysis may overcome them. For further assessing the effect of HC in clinical results a prospective controlled study, with a larger sample size could be performed, also comparing life-birth rates. Wider implications of the findings These results, alongside our previous findings (Valera et al. 2020, Albert et al. 2020), support that HC contributes to optimize embryo development and clinical results in undisturbed culture in TLS with single-step medium. To our knowledge, this is the largest study on the matter and the first performing multivariable analysis. Trial registration number Not applicable


2021 ◽  
Vol 36 (Supplement_1) ◽  
Author(s):  
S Mizumoto ◽  
H Watanabe ◽  
Y Nagao ◽  
K Tanaka ◽  
M Murakami ◽  
...  

Abstract Study question Does the addition of antioxidants for gamete preparation, insemination and embryo culture lead to differences in embryo development and clinical outcome Summary answer Using an antioxidant-containing media system for sperm preparation, insemination and embryo culture imparts significantly higher good-quality blastocyst rates and improved clinical outcome in elderly patients. What is known already A previous study showed that adding combined antioxidants for sequential embryo culture in conventional incubators (interrupted culture) improves embryo viability and clinical outcome, especially for elderly patients. Here we investigated the combined effect of three antioxidants Acetyl-L-Carnitine (10 µM), N-Acetyl-L-Cysteine (10 µM), and α-Lipoic Acid (5 µM) during sperm preparation, insemination, and time-lapse culture in a single step medium on human embryo development and clinical outcome. Study design, size, duration Prospective randomized single center study including 143 couples for IVF/ICSI between August 2018 and December 2019. Inclusion required at least eight cumulus-oocyte-complexes (COCs) after retrieval. Cycles involving PGT, split IVF/ICSI, and surgically retrieved sperm were excluded. Immediately after retrieval oocytes were randomly distributed to a study or control media system with or without antioxidants (Vitrolife). Similarly, ejaculates were split and prepared with and without antioxidants. Participants/materials, setting, methods Sibling oocytes were inseminated in the respective group with accordingly prepared sperm. Single step embryo culture was conducted in medium with (Gx-TL) and without (G-TL) antioxidants in the EmbryoScope+. Embryo quality and clinical outcome were assessed in relation to maternal age (<35/>35 years). Good-quality embryos on day 3 were defined as 8- to 10-cells with even cells and low fragmentation; good-quality blastocysts as > 3BB. Clinical outcome was assessed after single vitrified blastocyst transfer (SVBT). Main results and the role of chance From 143 participants (female age, 34.7±3.2 years), a total of 2424 COCs were collected; 1180 COCs/916 metaphase-II (MII) oocytes were allocated to Gx-TL media and 1244 COCs/981 MII oocytes to G-TL media. Age-related analysis in Gx-TL compared with G-TL in relation to allocated MII oocytes revealed a trend for higher fertilization rates in Gx-TL for both age groups (<35: 72.1% vs. 66.9%; >35: 70.7% vs. 64.9%, P < 0.1). Good-quality day 3 embryo development/MII oocytes was higher, albeit not significant, in the elderly patients in Gx-TL (<35: 35.9% vs. 34.4%; >35: 31.1% vs. 27.9%). Overall day 5/6 blastocyst rate was similar for both media (<35: 48.2% vs. 49.9%; >35: 42.3% vs. 39.5%). Day 5/6 GQB rate was comparable for younger patients (<35: 23.8% for Gx-TL vs. 26.0% for G-TL) but significantly higher in Gx-TL in elderly patients (>35: 20.7% vs. 14.4%; P < 0.05). A total of 200 SVBT were performed; 99 in the Gx-TL- and 101 in the G-TL-arm. We noted almost similar implantation and ongoing pregnancy rates between Gx-TL vs G-TL in the younger (<35) age group (50.0% vs. 55.4%; 50.0% vs. 55.6%) but higher albeit not significant rates for Gx-TL in older (>35) patients (44.1% vs. 33.3%; 44.1% vs. 33.3%). Limitations, reasons for caution In almost 95% of the cycles, oocytes were inseminated by ICSI; thus results may not equally apply for cycles with IVF. The use of a closed time-lapse system may have prevented from some environmental oxidative stress. Therefore results may come out different with a similar study using standard incubation. Wider implications of the findings: Supplementation of antioxidants to media for gamete isolation and preparation, as well as subsequent single step time-lapse culture may improve GQE/B rates and clinical outcomes in certain age groups, plausibly through the reduction of oxidative stress. Further studies in selected sub-groups (severe OAT syndrome / testicular cases) may be indicated. Trial registration number UMIN000034482


2021 ◽  
Vol 36 (Supplement_1) ◽  
Author(s):  
T A Vilori. Samochin ◽  
M A Valera ◽  
L Bori ◽  
F Meseguer ◽  
J M D Lo. Santos ◽  
...  

Abstract Study question Does culture in integrated time-lapse systems (TLS) improve embryo development and blastocyst quality compared to conventional benchtop incubators (CI), within the same IVF laboratory? Summary answer Under similar conditions, culture in TLS resulted in a significant increase in blastocyst rate, top quality blastocyst rate and proportion of biopsied embryos per treatment What is known already Integrated TLS have the potential of delivering a stable and undisturbed environment throughout the whole embryo culture, avoiding taking them out for assessment. However, there is still lack of quality evidence of the performance of these incubators compared to CI at supporting embryo culture until blastocyst stage. Studies abording this issue are still scarce, heterogeneous and have a small sample size. Although some authors have reported an improvement in embryo development and quality using TLS, global results are inconsistent. To our knowledge, the present study evaluates the effect of TLS on embryo quality on the largest sample size yet. Study design, size, duration Unicentric retrospective cohort study including 14248 ICSI treatments from 2016 to October 2020, with both autologous and donated oocytes. We compared blastocyst rate (BR) and proportion of top-quality blastocysts (TQB=Morphology ASEBIR score A) per treatment between those using TLS (N = 7500) and CI (N = 6748), and the proportion of embryos biopsied (EB) in cycles with pre-implantation genetic testing (PGT-A; N = 2642). We performed a sub-analysis in treatments using single-step culture medium (N-TLS=4398, N-CI=1140) in both types of incubators. Participants/materials, setting, methods Embryo cohorts were cultured until blastocyst stage in one of 3 TLS: EmbryoScope, EmbryoScope Plus (Vitrolife,) and Geri (Genea Biomedx), or in a CI (ASTEC). Embryo quality was assessed following ASEBIR morphological criteria. Culture protocols and media changed during the included time period. For that reason, we did a sub-study in the treatments performed since the implementation of Gems® (Genea Biomedx) single-step (SS) culture medium in all incubators. Statistical analysis was done using ANOVA tests. Main results and the role of chance Treatments were differently distributed and heterogeneous in terms of number of oocytes obtained per patient, so we stratified the analysis according to ovum origin and compared mean rates per cycle instead of total number of embryos per group. BR was statistically higher (P < 0,001) in the TLS group, in both autologous (62,98±29,37% vs 59,49±31,09% in CI) and oocyte donation treatments (69,25±22,07% vs 66,27±23,28% in CI). Proportion of TQB was also significantly higher in the TLS in both types of cycles (P < 0,001): 3,60±12,29% in TLS vs 2,27±9,71% in CI in autologous cycles, 8,68±15,31% in TLS vs 7,32±14,02% CI in ovum donation cycles. Results were corroborated in the SS media sub-study (P < 0,05): BR was 63,87±29,23% in TLS vs 57,53±30,61% in CI with autologous oocytes, and 70,76±21,63% in TLS vs 67,39±22,68% in CI with donated oocytes; TQB rates were 3,66±12,06% in TLS vs 2,05±9,26% in CI in autologous treatments and 8,81±15,21% in TLS vs 6,84±12,91% in CI in ovum donation treatments. Regarding PGT-A treatments, we found no significant difference in the biopsy rate in the total comparison, although the rate significantly increased in the TLS group since the implementation of single-step medium (52,36±24,69% in TLS vs 48,63±22,56% in CI; P = 0,007) Limitations, reasons for caution Not only culture conditions varied over time, but also the number of TLS in the laboratory, which increased lately. Hence, even though the most recent treatments included in the all-SS sub-study are more homogeneous in terms of culture conditions, they are unbalanced regarding the distribution among incubators. Wider implications of the findings: Our results demonstrate the superiority of TLS coupled with single-step culture media against traditional embryo culture systems at supporting embryo development. The optimal environment provided by TLS enhances embryo development until blastocyst stage as well as their quality, increasing the cumulative chances of getting a life-birth for each patien. Trial registration number Not applicable


2021 ◽  
Vol 36 (Supplement_1) ◽  
Author(s):  
D González-Abreu ◽  
E Mestre ◽  
M Escribá-Suárez ◽  
C Miret-Lucio ◽  
A García-Esteve ◽  
...  

Abstract Study question Can lab-related variables (media type, oil viscosity, microdroplet volume and culture dish design) modulate media evaporation and improve its stability during culture? Summary answer Using dishes with pre-defined wells, big volume microdroplets and high-viscosity mineral oil can help to reduce media evaporation and improve osmolality stability during embryo culture. What is known already Osmolality measures the number of solute particles present in a solution and is an important variable of a human embryo culture system. High ambient temperature and low humidity may induce evaporation in culture media, increasing its osmolality. In addition, recent tendencies in IVF laboratories, such as extending the embryo culture uninterruptedly until day 6/7 or the use of dry benchtop incubators, may intensify evaporation. Surpassing a 300mOsm/kg threshold can result deleterious for embryo development and impair clinical results. Different strategies (e.g. oil type/volume, dish type, micro-drop volume) have been proposed to reduce evaporation and stabilize osmolality during culture. Study design, size, duration Four variables were analyzed in their capacity to reduce media evaporation: type of culture medium, micro-droplet volume, oil viscosity and type of culture dish. Dishes were prepared with 5ml of oil and 50µl microdroplets (25µl were used for the comparison of micro-droplet volumes). Dishes were cultured in parallel in a dry benchtop incubator (AD–3100, Astec), and osmolality measured daily for seven days with a freezing point depression osmometer (Osmo1®, Advanced Instruments, accuracy ≤2mOsm/kg). Participants/materials, setting, methods The following comparison groups were analyzed: 1) Seven commercial single-step media with three differing initial osmolalities (approximately 260, 280 and >290mOsm/kg); 2) oil with high, medium or low viscosity; 3) 50 vs. 25µl microdroplets; 4) 35mm flat Petri dish vs. 35mm dish with defined wells. Temperature in the incubator was monitored continuously (T+Button, BrightSentinel), as well as room temperature and humidity (Octax Log&Guard, Vitrolife). All were stable at 37.3±0.05oC, 22.1±0.6 oC and 67.4±7.4%, respectively. Main results and the role of chance Evaporation occurred in all the studied groups, but its rate was modulated by various parameters. Culture dishes designed with pre-defined wells reduced evaporation when compared to regular Petri dishes (Increase 11.3mOsm/kg and increase 12.5mOsm/kg, respectively from day 0 to 7 (P = 0.007)). Similarly, oil viscosity had an impact in osmolality stability during culture, with an increase of 14.7mOsm/kg, 16.3mOsm/kg and 19.2mOsm/kg observed when using mineral oil with high, medium and low viscosity, respectively (P = 0.009). Finally, reducing the volume of the medium microdroplets from 50 to 25µl derived in higher evaporation rates, but without significant differences (Increase 14.7mOsm/kg and increase 15.8mOsm/kg, respectively (P = 0.325)). Different evaporation rates were observed between the seven studied culture media attending their three-differing initial osmolalities. Significant differences were observed for a media respect another three media with differing initial osmolality (P = 0.001, P = 0.01 and P = 0.015). Their initial osmolality had a direct correlation with the maximum osmolality reached at the end of culture. Thus, media with a high initial osmolality (>290mOsm/kg) resulted in hyperosmotic media above the recommended 300mOsm/kg threshold by the end of culture and, by contrast, the studied media with lower initial values were able to maintain osmolality below 300mOsm/kg for the whole duration of the culture. Limitations, reasons for caution While a clear effect was observed by the studied variables, other parameters, such as oil volume or dish preparation techniques, could also play a role in osmolality maintenance and could be studied in the future. Additionally, these findings could vary between different centers and should be validated in each laboratory. Wider implications of the findings: Osmolality has been shown to have a direct impact on embryo development, embryo quality and clinical outcomes. Carefully defining the consumables and methodologies used in the IVF laboratory will improve the stability of the culture system and, consequently, reduce the stress imparted to the embryos and gametes under culture. Trial registration number Not applicable


2021 ◽  
Vol 36 (Supplement_1) ◽  
Author(s):  
G Dionne ◽  
A J Watson ◽  
D H Betts ◽  
B A Rafea

Abstract Study question Our objective is determining whether supplementing embryo culture media with palmitic acid and/or oleic acid impacts Nrf2/Keap1 antioxidant response pathways during preimplantation mouse embryo development. Summary answer Supplementation of embryo culture media with palmitic acid increases cellular Nrf2 levels per embryo after 48-hour culture, while oleic acid reverses this effect. What is known already Obese women experience higher incidence of infertility than women with healthy BMIs. The obese reproductive tract environment supporting preimplantation embryo development is likely to include enhanced free fatty acid (FFA) levels and increased accumulation of reactive oxygen species. Exposure to palmitic acid (PA) in vitro significantly impairs mouse embryo development while increasing ER stress mRNAs. Oleic acid (OA) reverses these effects. To further define effects of FFA exposure, we are characterizing the influence of FFAs on the Nrf2–Keap1 pathway and its downstream antioxidant defense systems. We hypothesize that PA treatment induces Nrf2-Keap1 activity, while OA treatment alleviates pathway activity. Study design, size, duration Female CD–1 mice (4–6 weeks) were super-ovulated via intraperitoneal injections of PMSG, followed 48 hours later by hCG. Female mice were mated with male CD–1 mice (6–8 months) overnight. Females were euthanized using CO2 and two-cell embryos were collected by flushing oviducts. Two-cell embryos were placed into KSOMaa-based treatment groups: 1) BSA (control); 2) 100µM PA; 3) 100µM OA; 4) 100µM PA+OA, and cultured for 48 hours (37 °C; 5% O2, 5% CO2, 90% N2). Participants/materials, setting, methods After 48-hour embryo culture, developmental stages of all mouse embryos were recorded. Immunofluorescence analysis of Nrf2 and Keap1 localization was performed for embryo treatments (BSA, 100µM PA, 100µM OA & 100µM PA+OA) using rabbit polyclonal anti-Nrf2 antibody, with Rhodamine-Phalloidin and DAPI staining. Embryos were imaged using confocal microscopy and Nrf2-positive cells were counted using ImageJ. Nrf2 and Keap1 mRNA abundances were assessed after culture in each treatment condition using RT-qPCR and the delta-delta Ct method. Main results and the role of chance Inclusion of 100µM PA in embryo culture significantly decreased blastocyst development frequency from 70.06±16.38% in the BSA (control) group to 11.61±8.19% in the PA-treated group (p < 0.0001). Embryo culture with 100µM OA and 100µM PA+OA co-treatment did not significantly impair blastocyst development (OA: 61.59±8.07%, p = 0.4053; PA+OA: 63.53±7.63%, p = 0.6204). Embryo culture with PA treatment significantly increased the mean percentage of Nrf2-positive cells to 56.83±30.49% compared with 21.22±15.63% in the control group (p < 0.0001). Conversely, 100µM OA and 100µM PA+OA treatments did not significantly affect Nrf2-positive cell frequencies compared with the control group (OA: 33.28±21.83%, p = 0.1825; PA+OA: 34.84±12.66%, p = 0.0691). Immunofluorescence results show that treating embryos with 100µM PA for 48 hours results in increased levels of cellular Nrf2, while combining 100µM PA with 100µM OA reversed these effects. Preliminary qPCR analysis showed no significant differences in Nrf2 or Keap1 relative transcript abundance between any embryo treatment groups. Nrf2 and Keap1 mRNA levels were both higher after embryo culture with 100µM OA than all other culture groups (p = 0.6268; p = 0.3201). Notably, Keap1 relative transcript levels dropped to undetectable levels after culture with 100µM PA, which suggests an increase in Nrf2 activation.Limitations, reasons for caution: While immunofluorescence localization of Nrf2/Keap1 provides insight into how the proteins behave during preimplantation embryo development, confocal images cannot determine protein-protein interactions or activity levels. Similarly, transcript information from RT-qPCR analysis only provides information about Nrf2 and Keap1 at the transcript level. Nrf2 activity will be assessed via downstream targets. Wider implications of the findings: The Nrf2–Keap1 pathway coordinates numerous cellular defence mechanisms, and is implicated in various diseases, including cancer. Establishing an impact of free fatty acid exposure on Nrf2–Keap1 during preimplantation embryo development will provide valuable information regarding the effects of maternal obesity on outcomes for embryos produced from these patients. Trial registration number Not applicable


2021 ◽  
Vol 36 (Supplement_1) ◽  
Author(s):  
A Munuer. Puigvert ◽  
V. Montalv Pallès ◽  
J Mass. Hernáez ◽  
A García-Faura ◽  
B Marquè. López-Teijón ◽  
...  

Abstract Study question Have multinucleation and reverse cleavage any effect on embryo development and clinical outcomes on IVF treatments? Summary answer Embryos capable of repairing dysmorphisms and developing up to blastocyst stage keep intact their ability to become healthy babies. What is known already Time-lapse systems allow IVF laboratories to perform in-depth analysis of embryo development using the continuous monitoring tool. Some events that are impossible to detect with conventional morphologic evaluation, such as reverse cleavage or multinucleation, can be detected using time-lapse. Even though the low scientific evidence, the presence of these events is considered a negative factor when the embryo quality assessment is performed. However, it has been described the possibility that embryos have self-repair intrinsic methods. Study design, size, duration Retrospective study including data from 3,577 cycles with 21,274 embryos cultured until blastocyst stage using one-step culture media in time-lapse incubators (Embryoscope, Vitrolife) up to day 5/6 between 2014 and 2019. Participants/materials, setting, methods Three embryo groups were considered: Control group, embryos without multinucleation or reverse cleavage (CG; n = 16,897); Multinucleation group, embryos with at least one blastomere multinucleated on D + 2/3 (MNC; n = 3,879) and Reverse Cleavage group, embryos undergoing complete fusion of two blastomeres on D + 2/3 (RC; n = 498). Single embryo transfer was performed on blastocyst stage. Clinical outcome rates were compared between groups and analyzed by Chi-square test. Main results and the role of chance As published by other groups, the 2.3% of our embryos showed at least one reverse cleavage event and we observed multinucleation in the 18.2% of the embryos. Blastocyst rate of dysmorphism groups was significantly lower (p < 0.05) than Control group (MNC=20.0%; RC = 27.7%; CG = 58.0%). Once transferred, MNC and RC evolutive embryos showed significantly lower pregnancy (MNC=47.9%; RC = 46.8%; CG = 60.8%; p < 0.05) and clinical pregnancy rates (MNC=39.4%; RC = 40.4% CG = 50.6%; p < 0.05) than the Control group (p < 0.05). However, during the post-implantational development the negative effect of dysmorphisms disappears, reaching values of live birth rate comparable to the Control group (MNC=28.3%; RC = 31.9% CG = 33.8%; p = 0.17). These results prove the importance of blastocyst culture and the inherent capability of the embryos to overcome some abnormal dynamics as multinucleation and reverse cleavage. Thus, these embryos showing the poor-prognosis events can be considered for transfer or vitrify. Limitations, reasons for caution There is a wide difference on sample size between groups despite the fact that the statistical analysis considers that into account. There are some ongoing pregnancies in all groups. Wider implications of the findings: When analyzing the development of embryos undergoing reverse cleavage and multinucleation, we hypothesize that these embryos could be showing a self-correction mechanism for some type of error detected. Embryos capable of repairing and developing up to blastocyst stage keep intact their ability to become healthy babies. Trial registration number Not applicable


2013 ◽  
Vol 100 (3) ◽  
pp. S246
Author(s):  
M.A. Stout ◽  
V.A. Cholewczynski ◽  
S.G. Shukry ◽  
G.S. Karman ◽  
J.R. Graham ◽  
...  

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