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Impact of oxygen concentration on embryo development, embryo morphology and morphokinetics
Women’s Health, Division of Reproductive Endocrinology, Albert Einstein College of Medicine and Montefiore Medical Center, Bronx, NY. OBJECTIVE: To evaluate if diagnosis is predictive of inner cell mass (ICM) and/or trophectoderm (TE) scores and whether TE score correlates to implantation rate (IR). DESIGN: Retrospective. MATERIALS AND METHODS: Women undergoing a fresh, autologous IVF or ICSI cycle from 1/2007–12/2010 were included. Fertility diagnoses were tubal factor, male factor, PCOS, unexplained, endometriosis, diminished ovarian reserve, uterine and other. Analysis was performed for patients whose cultured embryos were transferred on day 5. Observations of blastocyst expansion, ICM and TE were based on Veeck et al. 2004. Composite TE grades were derived for all blastocysts (n¼307) per transfer (n¼149). Statistical methods included Pearson’s Chi Square, Fishers Exact Analysis, ANOVA and multivariate logistic regression. p <0.05 was considered significant. RESULTS: 1727 embryos from 142 patients were included. Patients had an overall blastulation rate of 42.8%. Blastocysts from women with unexplained infertility were significantly more likely to have poor TE score vs good/fair TE score (p¼0.03). Similarly, blastocysts from women with unexplained infertility or endometriosis were significantly more likely to have a poor ICM score vs good/fair ICM score (p¼0.02 and p¼0.04, respectively). After controlling for age, maximum FSH, number of oocytes retrieved and mature, unexplained infertility was a significant and independent negative predictor of TE score (OR¼0.66, CI: 0.47-0.94, p¼0.02). Stratifying IR by diagnosis, we did not find a relationship of composite TE score to IR (p¼0.79). In patients with unexplained infertility, there was a higher composite TE score, indicating poorer TE grades, when compared to patients with all other diagnoses. CONCLUSION: TE score has recently been shown to predict pregnancy outcome. We demonstrate that unexplained infertility predicts poor TE grade but not IR. Additional powered analysis will be conducted to establish whether poor TE grades are associated with live birth rates in unexplained infertility patients. P-321 Tuesday, October 15, 2013 IMPACT OF OXYGEN CONCENTRATION ON EMBRYO DEVELOPMENT, EMBRYO MORPHOLOGY AND MORPHOKINETICS. N. Zaninovic, J. Goldschlag, H. Yin, Z. Ye, R. Clarke, Z. Rosenwaks. CRM, Reproductive Medicine, Weill Cornell Medical College of Cornell University, New York, NY. OBJECTIVE: To compare embryo development, morphology and morphokinetics of sibling oocytes incubated in the EmbryoScopeÒ and standard incubators under different oxygen concentrations and culture time lines. DESIGN: Clinical study. MATERIALS AND METHODS: Study 1: Embryo development, morphology and morphokinetics of zygotes (n¼1799, 144 patients) incubated in standard incubator (S, 20% O2), E-EmbryoScope (20% O2) or 3E-EmbryoScope (5% O2) were compared. 1. S vs. E (n¼77); 2: S vs.3E (n¼35); 3: 3E vs. E (n¼28) and 4: S vs. E vs. 3E (n¼4). Study 2: Development of 565 oocytes or zygotes (77 ICSI patients) incubated in the Embryoscope 3E (5% O2) immediately after ICSI (day 0) vs. the day after fertilization check (S-3E, 20% O2; 16-20 hours hrs. in 20% O2). RESULTS: Study 1. Improved embryo development was observed after incubation with the EmbryoScope compared to standard incubators. More embryos were transferred and more blastocysts (BL) were frozen in the 3E group vs. E or S (p<0.05). Group 1 and 2: E- or 3E-incubated embryos superior to S-incubated embryos for all parameters. 3E exceeded E in all parameters in group 3 and E and S in group 4 .Morphokinetic analysis between E and 3E (impact of reduced oxygen) revealed a significant shift in third cleavage where 3E embryos expressed earlier 5-cell onset than E, this was phenomena was subsequently reproduced in 6-8 cells to BL. Study 2. Embryos cultured entirely in 5% O2 (3E) had significantly better embryo development than the transient 20% O2 exposure (S-3E) group. More embryos were transferred and more BL were frozen from the former (p<0.05). Morphokinetic analysis revealed embryos reaching 4 to 8 cells and morula stage faster in 3E vs. S-3E. CONCLUSION: EmbryoScope is superior to the standard incubator. Reduced oxygen improved embryo development. Even transient exposure of the embryos to 20% O2 slowed embryo development. These findings in combination with improved embryo selection criteria suggests that the EmbryoScope and 5% O2 is superior to standard incubators. Supported by: Institutional.
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ASRM Abstracts
P-322 Tuesday, October 15, 2013 THE PREDICTIVE VALUE OF ANTI-MULLERIAN HORMONE ON EMBRYO QUALITY FOLLOWING IN VITRO FERTILIZATION. O. Alag€oz,a R. Attar,b G. Yildirim,b P. Cenksoy,b M. Yesiladali,b C. Ficicioglu.b aDepartment of Histology and Embryology, Yeditepe University Hospital, Istanbul, Turkey; bDepartment of Obstetrics and Gynecology, Yeditepe University Hospital, Istanbul, Turkey. OBJECTIVE: High AMH levels on Day 3 have been shown to be corelated with high numbers of mature oocytes, resulting in more embryos and higher clinical pregnancy rate, previously. As oocyte quantity and oocyte quality affect embryo quality, AMH might also correlate with embryo quality. In this study we aimed to evaluate the association between basal serum levels of anti-Mullerian hormone (AMH) and embryo quality following in vitro fertilization (IVF). DESIGN: This is a retrospective study. MATERIALS AND METHODS: 200 women who underwent in vitro fertilisation (IVF) / intracytoplasmic sperm injection (ICSI) cycles at the IVF center of Yeditepe University Hospital were included in this study. Day three basal AMH serum levels and embryo quality which were transferred to the patients were evaluated. RESULTS: The mean day three AMH levels were 2.04 0.21 ng/ mL(SEM), 1.280.12 ng/mL(SEM), 1.530.12 ng/mL(SEM) for Grade I, Grade II and Grade III embryos, respectively. We observed that AMH levels were significantly higher in women with grade I embryos (p¼0.01). We used Spearman correlation analysis to evaluate the association between AMH and embryo quality. Spearman correlation analysis indicated that there was a negative correlation between the AMH level and Grade of the embryos which meant that as the AMH levels decreased embryo Grade increased. Higher levels of AMH were associated with Grade I embryos and the difference was statistically significant (p¼0.001). CONCLUSION: Good quality embryos are identified on the basis of morphological findings. It has been suggested that AMH might reflect embryo morphology. However there are conflicting result about AMH and its predictive value for embryo morphology in the literature. Some investigators state that AMH is a predictive indicator of the ovarian reserve, in terms of quantity but not in terms of quality. In this study, we showed that serum AMH levels were positively correlated with good quality embryos. It may be used as a marker of IVF outcome in the future. P-323 Tuesday, October 15, 2013 TO WHAT EXTENT HUMAN EMBRYO VIABILITY IS CONDITIONED BY CLEAVAGE TIMES? A TIME-LAPSE-ANALYSIS ON 9530 EMBRYOS. J. Herrero, I. Rubio, A. Tejera, C. Vidal, J. L. Romero, M. Meseguer. Instituto Universitario IVI Valencia, Valencia, Spain. OBJECTIVE: Analyze variations in specific time-points of embryo development according to viability. DESIGN: Retrospective study including 9530 human embryos from 1806 first or second ICSI cycles between September 2010 and July 2012 at IVI Valencia, Spain. MATERIALS AND METHODS: A time-lapse instrument automatically acquired images of the embryos every 15 minutes. Cleavages times from first cleavage on (t2 and forward) and variables as the length of second cell cycle (cc2 ¼ t3-t2) or synchrony from 2 to 4 cells (s2 ¼ t4-t3) were mesured. Viability was represented clustering embryos into three experimental groups: ‘‘regular divisions’’ excluded direct cleaving embryos from 1 to 3 or 2 to 5 cells; ‘‘viable 8 cell’’ and ‘‘viable blastocyst’’ included only embryos capable to reach these stages. Medians and graphical distribution of the times were analyzed and compared between the groups. RESULTS: In ‘‘viable 8 cells’’ medians were lower for t2, t3, t5, cc2, cc3, s2 and s3 compared with regular divisions (p<0.05) indicating the presence of slower embryos in the starting population that were not capable of reaching the 8-cell stage. However in ‘‘viable blastocyst’’ medians were higher for t5, t8, tM, cc3 and s2 compared with viable 8 cells (p<0,05) suggesting that rather slower embryos take advantage of reaching the blastocyst stage while fast ones are penalized.The highest dispersion of data occurred in ‘‘regular divisions’’ being the lowest in ‘‘viable blastocyst’’ revealing that many of the delayed cleaving embryos did not reach the blastocyst stage. Dispersion of values in ‘‘viable blastocyst’’ was lower in early variables suggesting that flexibility in the ranges of time required for reaching the blastocyst stage increases as development progresses.
Vol. 100, No. 3, Supplement, September 2013
S240
ASRM Abstracts
P-322 Tuesday, October 15, 2013 THE PREDICTIVE VALUE OF ANTI-MULLERIAN HORMONE ON EMBRYO QUALITY FOLLOWING IN VITRO FERTILIZATION. O. Alag€oz,a R. Attar,b G. Yildirim,b P. Cenksoy,b M. Yesiladali,b C. Ficicioglu.b aDepartment of Histology and Embryology, Yeditepe University Hospital, Istanbul, Turkey; bDepartment of Obstetrics and Gynecology, Yeditepe University Hospital, Istanbul, Turkey. OBJECTIVE: High AMH levels on Day 3 have been shown to be corelated with high numbers of mature oocytes, resulting in more embryos and higher clinical pregnancy rate, previously. As oocyte quantity and oocyte quality affect embryo quality, AMH might also correlate with embryo quality. In this study we aimed to evaluate the association between basal serum levels of anti-Mullerian hormone (AMH) and embryo quality following in vitro fertilization (IVF). DESIGN: This is a retrospective study. MATERIALS AND METHODS: 200 women who underwent in vitro fertilisation (IVF) / intracytoplasmic sperm injection (ICSI) cycles at the IVF center of Yeditepe University Hospital were included in this study. Day three basal AMH serum levels and embryo quality which were transferred to the patients were evaluated. RESULTS: The mean day three AMH levels were 2.04 0.21 ng/ mL(SEM), 1.280.12 ng/mL(SEM), 1.530.12 ng/mL(SEM) for Grade I, Grade II and Grade III embryos, respectively. We observed that AMH levels were significantly higher in women with grade I embryos (p¼0.01). We used Spearman correlation analysis to evaluate the association between AMH and embryo quality. Spearman correlation analysis indicated that there was a negative correlation between the AMH level and Grade of the embryos which meant that as the AMH levels decreased embryo Grade increased. Higher levels of AMH were associated with Grade I embryos and the difference was statistically significant (p¼0.001). CONCLUSION: Good quality embryos are identified on the basis of morphological findings. It has been suggested that AMH might reflect embryo morphology. However there are conflicting result about AMH and its predictive value for embryo morphology in the literature. Some investigators state that AMH is a predictive indicator of the ovarian reserve, in terms of quantity but not in terms of quality. In this study, we showed that serum AMH levels were positively correlated with good quality embryos. It may be used as a marker of IVF outcome in the future. P-323 Tuesday, October 15, 2013 TO WHAT EXTENT HUMAN EMBRYO VIABILITY IS CONDITIONED BY CLEAVAGE TIMES? A TIME-LAPSE-ANALYSIS ON 9530 EMBRYOS. J. Herrero, I. Rubio, A. Tejera, C. Vidal, J. L. Romero, M. Meseguer. Instituto Universitario IVI Valencia, Valencia, Spain. OBJECTIVE: Analyze variations in specific time-points of embryo development according to viability. DESIGN: Retrospective study including 9530 human embryos from 1806 first or second ICSI cycles between September 2010 and July 2012 at IVI Valencia, Spain. MATERIALS AND METHODS: A time-lapse instrument automatically acquired images of the embryos every 15 minutes. Cleavages times from first cleavage on (t2 and forward) and variables as the length of second cell cycle (cc2 ¼ t3-t2) or synchrony from 2 to 4 cells (s2 ¼ t4-t3) were mesured. Viability was represented clustering embryos into three experimental groups: ‘‘regular divisions’’ excluded direct cleaving embryos from 1 to 3 or 2 to 5 cells; ‘‘viable 8 cell’’ and ‘‘viable blastocyst’’ included only embryos capable to reach these stages. Medians and graphical distribution of the times were analyzed and compared between the groups. RESULTS: In ‘‘viable 8 cells’’ medians were lower for t2, t3, t5, cc2, cc3, s2 and s3 compared with regular divisions (p<0.05) indicating the presence of slower embryos in the starting population that were not capable of reaching the 8-cell stage. However in ‘‘viable blastocyst’’ medians were higher for t5, t8, tM, cc3 and s2 compared with viable 8 cells (p<0,05) suggesting that rather slower embryos take advantage of reaching the blastocyst stage while fast ones are penalized.The highest dispersion of data occurred in ‘‘regular divisions’’ being the lowest in ‘‘viable blastocyst’’ revealing that many of the delayed cleaving embryos did not reach the blastocyst stage. Dispersion of values in ‘‘viable blastocyst’’ was lower in early variables suggesting that flexibility in the ranges of time required for reaching the blastocyst stage increases as development progresses.
Vol. 100, No. 3, Supplement, September 2013