O-214 Undisturbed embryo culture under High Humidity atmosphere in a time-lapse system increases pregnancy rates

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):  
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):  
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):  
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


Author(s):  
Mariano Mascarenhas ◽  
Sarah J Owen ◽  
Emily French ◽  
Karen Thompson ◽  
Adam H Balen

Objective To compare the cumulative live birth rate per egg retrieval between time lapse imaging (TLI) incubators and standard culture (SC) incubators both using a single-step culture medium Design Retrospective cohort study Setting A tertiary level fertility-centre Population Women undergoing an IVF cycle between November 2015 and December 2017 Methods Comparison was done between 1219 IVF cycles using TLI and 1039 cycles using SC after accounting for confounding factors such as age and number of oocytes retrieved. Main outcome measure Cumulative live birth rate per egg retrieval Results The live birth rate per egg retrieval following fresh embryo transfer was noted to be higher for TLI cycles (TLI 39.87% vs SC 38.02%, aOR 1.20, 95% CI 1.01 to 1.44). More embryos were available for cryopreservation in the TLI arm (MD 0.08 embryos, 95% CI 0.10 to 0.41). The live birth rate per frozen embryo transfer was not significantly different. The cumulative live birth rate per egg retrieval was significantly higher in the TLI arm (TLI 50.29% vs SC 46.78%, aOR 1.24, 95% CI 1.04 to 1.48) Conclusions With the use of single step medium, there appears to be a greater benefit of TLI through a reduced interruption in embryo culture conditions, resulting in a higher number of embryos available for cryostorage which in turn appears to improve the cumulative live birth rate. Funding No funding was obtained for this study Keywords Time lapse imaging, cumulative live birth rate, single step culture medium, embryo utilization rate


2017 ◽  
Vol 107 (2) ◽  
pp. 405-412 ◽  
Author(s):  
Mohamed Fawzy ◽  
Mohamed Sabry ◽  
Mohamed Nour ◽  
Mohamed Y. Abdelrahman ◽  
Eman Roshdy ◽  
...  

2021 ◽  
Vol 116 (3) ◽  
pp. e181-e182
Author(s):  
Rebecca Holmes ◽  
Rebecca Kile ◽  
Heather Rogers ◽  
Ye Yuan ◽  
William B. Schoolcraft ◽  
...  

2020 ◽  
pp. 47-50
Author(s):  
N. V. Saraeva ◽  
N. V. Spiridonova ◽  
M. T. Tugushev ◽  
O. V. Shurygina ◽  
A. I. Sinitsyna

In order to increase the pregnancy rate in the assisted reproductive technology, the selection of one embryo with the highest implantation potential it is very important. Time-lapse microscopy (TLM) is a tool for selecting quality embryos for transfer. This study aimed to assess the benefits of single-embryo transfer of autologous oocytes performed on day 5 of embryo incubation in a TLM-equipped system in IVF and ICSI programs. Single-embryo transfer following incubation in a TLM-equipped incubator was performed in 282 patients, who formed the main group; the control group consisted of 461 patients undergoing single-embryo transfer following a traditional culture and embryo selection procedure. We assessed the quality of transferred embryos, the rates of clinical pregnancy and delivery. The groups did not differ in the ratio of IVF and ICSI cycles, average age, and infertility factor. The proportion of excellent quality embryos for transfer was 77.0% in the main group and 65.1% in the control group (p = 0.001). In the subgroup with receiving eight and less oocytes we noted the tendency of receiving more quality embryos in the main group (р = 0.052). In the subgroup of nine and more oocytes the quality of the transferred embryos did not differ between two groups. The clinical pregnancy rate was 60.2% in the main group and 52.9% in the control group (p = 0.057). The delivery rate was 45.0% in the main group and 39.9% in the control group (p > 0.050).


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