scholarly journals SINGLE-STEP VS SEQUENTIAL MEDIA: UNBIASED COMPARISON OF CLINICAL OUTCOME BETWEEN TWO INDIVIDUAL EMBRYO CULTURE STRATEGIES IN A TIME-LAPSE INCUBATOR

2021 ◽  
Vol 116 (3) ◽  
pp. e5
Author(s):  
María de los Ángeles Valera ◽  
Lorena Bori ◽  
Fernando Meseguer ◽  
Elena Paya ◽  
Ma José de los Santos ◽  
...  
Keyword(s):  
Clinical Outcome ◽  
Embryo Culture ◽  
Time Lapse ◽  
Single Step ◽  
Culture Strategies

P–221 Prospective randomized sibling study on gamete preparation, insemination and subsequent culture of human oocytes in a time-lapse system using media systems with and without antioxidants

2021 ◽  
Vol 36 (Supplement_1) ◽  
Author(s):  
S Mizumoto ◽  
H Watanabe ◽  
Y Nagao ◽  
K Tanaka ◽  
M Murakami ◽  
...  
Keyword(s):  
Oxidative Stress ◽  
Clinical Outcome ◽  
Elderly Patients ◽  
Embryo Development ◽  
Embryo Culture ◽  
Age Groups ◽  
Time Lapse ◽  
Single Step ◽  
Sperm Preparation ◽  
Media System

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

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 ◽  
...  
Keyword(s):  
Logistic Regression ◽  
Embryo Development ◽  
Embryo Culture ◽  
Culture Media ◽  
Time Lapse ◽  
Clinical Results ◽  
Single Step ◽  
Ongoing Pregnancy ◽  
Media Change ◽  
Culture Strategy

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

P–144 Undisturbed culture in time-lapse systems improves embryo development and quality

2021 ◽  
Vol 36 (Supplement_1) ◽  
Author(s):  
T A Vilori. Samochin ◽  
M A Valera ◽  
L Bori ◽  
F Meseguer ◽  
J M D Lo. Santos ◽  
...  
Keyword(s):  
Embryo Development ◽  
Embryo Culture ◽  
Culture Medium ◽  
Embryo Quality ◽  
Time Lapse ◽  
Blastocyst Stage ◽  
Culture Conditions ◽  
Single Step ◽  
Ovum Donation ◽  
Blastocyst Rate

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

P–561 Male and female blastocyst: any difference other than the sex?

2021 ◽  
Vol 36 (Supplement_1) ◽  
Author(s):  
B Carrasc. Canal ◽  
M C Pons ◽  
M Parriego ◽  
M Boada ◽  
S García ◽  
...  
Keyword(s):  
Embryo Transfer ◽  
Sperm Quality ◽  
Time Lapse ◽  
Blastocyst Stage ◽  
Study Data ◽  
Single Step ◽  
Chromosomal Anomalies ◽  
Low Oxygen ◽  
Male And Female ◽  
Normal Copy Number

Abstract Study question Is there any imbalance in the sex ratio (SR) and in the aneuploidy rate of male and female human blastocysts from a PGT-A programme? Summary answer Although SR in human blastocysts is significantly male-biased, more aneuploidies are observed among male blastocysts, resulting in comparable euploid male and female embryos available. What is known already More boys than girls are born worldwide, meaning that the SR at birth is biased towards males. Differences in the SR of children born after ART have been also reported. Factors such as the insemination technique or the day of embryo transfer have been shown to be related to the SR at birth, but whether the SR is shifted during the preimplantation and/or postimplantation development remains unknown. Study design, size, duration: Embryos from patients undergoing 921 PGT-A cycles from September 2017 to February 2020 were included in the study. Data from the chromosomal constitution of 2637 biopsied blastocysts was retrospectively analysed. Participants/materials, setting, methods Embryos were cultured in time-lapse incubators with low oxygen tension (5%) (Embryoscope®; Geri®) using single-step medium (Global®, LifeGlobal®; GTL™, Vitrolife). Blastocyst biopsy was performed between D5-D7 followed by immediate vitrification (Cryotop®, Kitazato). Trophectoderm samples were analysed by NGS. Embryos were categorized as euploid, aneuploid or mosaic. Embryos were called as mosaic when the deviation from the normal copy number was ≥30% and <70%. Main results and the role of chance Overall biopsies from 2637 blastocysts were analysed, 1320 on day 5 (50.1%), 1169 on day 6 (44.3%) and 148 on day 7 (5.6%). Sex distribution among the embryos analysed was skewed in favor of male sex with 1401 diagnosed as male (53.1%) and 1236 were female (46.9%), [OR (95%CI):1.13(1.05–1.22)]. As a consequence of this biased SR, more male embryos reached the blastocyst stage and were biopsied both on day 5/6 (708/1320, 53.6% on day 5 and 619/1169, 53% on day 6). Embryos biopsied on day 7 were balanced between sexes with 50% being male and 50% being female. Following biopsy and PGT-A, 1086 (41.2%) of the embryos were classified as euploid, 1349 (51.16%) as aneuploid, and 202 (7.7%) as mosaic embryos. More chromosomal anomalies were observed among male blastocysts when compared to the female ones, 738 (52.7%) vs 611 (49.4%). Similarly, mosaicism was more frequents in male as compared with female blastocysts, 123 (8.8%) vs 79 (6.4%). (P = 0.000). As more aneuploidies are observed among male blastocysts, the final number of available euploid blastocysts for embryo transfer was comparable between sexes (540 male/546 female), [OR (95%CI): 0.99 (0.87–1.11)]. Limitations, reasons for caution This is a retrospective study. Only embryos at the blastocyst stage have been analyzed. Potential confounding factors such as sperm quality or the female age have not been analyzed. No data regarding the SR at birth have been analyzed in these study. Wider implications of the findings: In our study, more male embryos develop to the blastocyst when compared to female ones. It can be hypothesized that female embryos can be more affected by an early arrest at cleavage stages. SR at birth would be expected to be similar as more aneuploidy is observed in male embryos. Trial registration number Not applicable

EMBRYO CULTURE WITH SEQUENTIAL VS SINGLE STEP MEDIA: EFFECT ON EUPLOID RATES?

2020 ◽  
Vol 114 (3) ◽  
pp. e133
Author(s):  
Krishna Mantravadi ◽  
Gayathri Gedela ◽  
Durga Gedela Rao ◽  
Sandeep Karunakaran
Keyword(s):  
Embryo Culture ◽  
Single Step ◽  
Media Effect

Transfer of embryos following culture in continuous single-step or sequential-step embryo culture media

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

scholarly journals Embryo incubation and assessment by time-lapse system versus conventional incubators in Chinese women with diminished ovarian re serve undergoing IVF/ICSI : a study protocol for a randomized controlled trial

2020 ◽  
Author(s):  
Miaoxin Chen ◽  
Yuanyuan Wu ◽  
Xin Huang ◽  
Wentao Li ◽  
Chunyan Sun ◽  
...  
Keyword(s):  
Clinical Trial ◽  
Live Birth ◽  
Embryo Culture ◽  
Birth Rate ◽  
Live Birth Rate ◽  
Time Lapse ◽  
Absolute Difference ◽  
Cumulative Live Birth Rate ◽  
Randomized Controlled ◽  
Cumulative Live Birth

Abstract Background: Time-lapse imaging system (TLS) is a newly developed non-invasive embryo assessment system. Compared with conventional incubators, it provides stable culture condition and consistent observation of embryo development, thereby improving embryo quality and selection. In theory, these benefits could improve clinical outcomes of in vitro fertilization (IVF) or intracytoplasmic sperm injection (ICSI). Although this system has been routinely used in many IVF centers globally, it remains unclear if the TLS results in higher cumulative live birth rate and high-quality evidence is warranted. The purpose of this study is to compare the effectiveness of the TLS with conventional incubators in infertile diminished ovarian reserve (DOR) patients. Methods: This study is a double-blind randomized controlled clinical trial (1:1 treatment ratio of TLS vs. conventional incubator). A total of 730 DOR patients undergoing the first or second cycle of IVF or ICSI will be enrolled and randomized into two parallel groups. Participants in group A will undergo embryo culture and selection in the TLS, and participants in group B will undergo embryo culture in the conventional incubators and embryo selection by the morphological characteristics. The primary outcome is the cumulative live birth rate of the trial IVF/ICSI cycle within 12 months after randomization. This study is powered to detect an absolute difference of 10% (35% vs 25%) at the significance level of 0.05 and 80% statistical power based on a two-sided test. Discussion: The results of this study will provide evidence for the efficacy and safety of time-lapse system compared with conventional incubators in patients with DOR undergoing IVF/ICSI. Trial registration: Chinese Clinical Trial Registry, ChiCTR1900027746. Registered on 24 Nov 2019.

scholarly journals Impact of time-lapse imaging incubators with single-step culture medium on cumulative live birth rate in IVF cycles: a retrospective cohort study

2021 ◽  
Author(s):  
Mariano Mascarenhas ◽  
Sarah J Owen ◽  
Emily French ◽  
Karen Thompson ◽  
Adam H Balen
Keyword(s):  
Live Birth ◽  
Retrospective Cohort ◽  
Birth Rate ◽  
Culture Medium ◽  
Live Birth Rate ◽  
Time Lapse ◽  
Single Step ◽  
Cumulative Live Birth Rate ◽  
Cumulative Live Birth ◽  
Time Lapse Imaging

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

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