- Email: [email protected]
Comparison of vitrification devices for human embryos between open and closed system
CONCLUSION: Aneuploidy rates in egg donors vary significantly per center. The differences were not attributable to cohort size (blastocysts biopsied). This is the first study to investigate the question of center-dependent aneuploidy controlling for indication and patient population by taking into account only egg donor cycles. The reasons for the observed differences between centers can only be speculated but include any of the factors that are known to affect oocyte and embryo development, such as hormonal stimulation regimes, laboratory conditions, gamete and embryo manipulation methods, or environmental differences. Age is not considered to play a role here since young age is a pre-requisite to anonymous egg donation. O-355 Wednesday, October 22, 2014 05:30 PM DEVELOPMENT OF A TARGETED SINGLE MOLECULE SEQUENCING PANEL FOR OVARIAN HYPERSTIMULATION SYNDROME. T. J. O’Brien,a A. F. Harralson,b D. Frankfurter,c P. Gindoff,d F. Orkunoglu-Suer.e aGenetics, Quest Diagnostics/Athena Diagnostics, Worcester, MA; bPharmacogenomics, Shenandoah University, Ashburn, VA; cObstetrics and Gynecology, The George Washington University, Washington, DC; dPharmacology and Physiology, The George Washington University, Washington, DC. OBJECTIVE: The objective of this study was to identify potential genetic biomarkers for ovarian hyperstimulation syndrome (OHSS) using targeted single molecule sequencing (T-SMS). DESIGN: This was a retrospective, observational study. MATERIALS AND METHODS: DNA was isolated from patient blood samples. A T-SMS panel for genes implicated in ovarian response to controlled ovarian hyperstimulation was developed. SMS was carried out using the Pacific Biosciences single molecule, real-time, DNA Sequencing System. Primary data analyses, alignment and filtering utilized the Pacific Biosciences SMRT portal. Secondary analyses was conducted using the Genome Analysis Toolkit for SNP discovery and wANNOVAR for functional analysis of exonic variants. Filtered functional variants were further validated using conventional Sanger sequencing. RESULTS: Target enrichment using droplet-based, multiplex polymerase chain reaction generated amplicons averaging 1 kb fragment size from 44 target loci (99.8% unique base-pair coverage, 3.18 Mb per sample). SMS produced an average raw read length of 1178 nucleotides (nt) with 5% of the amplicons >6000 nt. After filtering with circular consensus (CCS) reads, the mean read length was 3200 nt (97% CCS accuracy). A total of 46 exonic variants were initially identified with 6 observed in OHSS, 24 in non-OHSS and 16 found in both. All variants were validated by Sanger DNA sequencing. CONCLUSION: These results offer promise of identifying genetic biomarkers for OHSS risk in controlled ovarian hyperstimulation patients. To the best of our knowledge, this is the first report utilizing emulsion PCR and T-SMS for long reads using human DNA samples. Supported by: This study was funded by award Number UL1RR031988 from the National Center for Research Resources. The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Center for Research Resources or the National Institutes of Health.
CRYOPRESERVATION AND FROZEN EMBRYO TRANSFER II O-356 Wednesday, October 22, 2014 03:45 PM EFFICACY AND EFFICIENCY OF THE ‘‘UNIVERSAL WARMING PROTOCOL’’: MULTICENTER RANDOMIZED CONTROLLED STUDY ON HUMAN SLOW FROZEN OOCYTES. L. Parmegiani,a C. Garello,b A. Revelli,b L. Criscuoli,c S. Dabizzi,c R. Gualtieri,d R. Talevi,d M. Filicori.a aReproductive Medicine Unit, GynePro Medical Centers, Bologna, BO, Italy; bLivet Clinic, Turin, TO, Italy; cCentre of Reproductive Medicine,, Careggi University Hospital, Florence, FI, Italy; d Dipartimento di Biologia, Universita di Napoli ‘‘Federico II’’, Naples, Italy. OBJECTIVE: Human reproductive cells are cryopreserved via slow freezing (SF) or vitrification (VIT). In a previously published pilot-
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ASRM Abstracts
study, it has already been demonstrated that it is possible to warm SF oocytes by using warming solution for VIT and also to increase their survival rates (1). The aim of the present study was to confirm the efficacy of the ‘‘rapid warming protocol’’ for SF oocytes in a multicenter trial and to compare its efficiency with the standard rapid thawing protocols. DESIGN: Three Assisted Reproductive (AR) centers in Italy were involved in this study. This was a prospective study on 414 oocytes randomized for conventional rapid thawing (RT) via two ready-to-use thawing media (Oocyte Thawing-Kit Origio–group A; CSC Thawing Medium Kit – group B), or rapid warming (RW) via VIT warming solutions (Vitrification Warming-Kit Sage – group C). MATERIALS AND METHODS: Primary endpoint was the morphological assessment of survival at 2 hours. Some of the surviving oocytes were divided into 3 sub-groups: i) parthenogenetically activated and observed by time-lapse microscopy ii) fixed and observed by confocal microscopy, iii) evaluated by polarized light for the presence of the meiotic spindle. Secondary endpoints were parthenogenetic activation and development, the assessment of the markers of oocyte preservation by confocal microscopy and the spindle presence by polarized light. RESULTS: Survival rate was 80.7% (71/88) in group A, 60.4% (125/ 207) in group B and 91.6 (109/119) in group C. Survival rate was significantly higher in group C (P¼0.036 vs group A and P<0.001 vs group B), in which the SF oocytes were warmed via VIT warming solutions. Survival rate was also significantly higher in group A vs group B (P¼0.001) Parthenogenetic activation and meiotic spindle presence were not significantly different in the three groups. At the time of writing this abstract the evaluation by confocal microscopy is still in progress. CONCLUSION: The findings of this study seem to confirm that is possible to optimize costs and increase the survival rate by using the same warming protocol for both SF and VIT oocytes. This ‘‘universal warming protocol’’ is potentially applicable to all kind of slow frozen reproductive cells, such as zygotes, cleavage stage embryos and blastocysts. Supported by: Thawing and warming solution were kindly supplied by Origio. O-357 Wednesday, October 22, 2014 04:00 PM COMPARISON OF VITRIFICATION DEVICES FOR HUMAN EMBRYOS BETWEEN OPEN AND CLOSED SYSTEM. S. Mizuno,a A. Ohgaki,a H. Matsumoto,a A. Fukuda,a Y. Morimoto.b aIVF Osaka Clinic, Higashi-Osaka, Osaka, Japan; bIVF Namba Clinic, Nishi-ku Osaka, Osaka, Japan. OBJECTIVE: Vitrification is a freezing method that is applied in almost every IVF clinic in Japan. In our clinic, human embryos are vitrified by Cryotop method in which the solution containing embryos is directly exposed to liquid nitrogen. This method does not completely eliminate the risk of cross-contamination during their storage. Therefore, the closed vitrification device, Rapid-i, has been developed to solve this problem. In the present study, survival rate and subsequent development of warmed embryos were compared between Cryotop and Rapid-i, to evaluate if embryos can be safely cryopreserved by Rapid-i without sacrificing their potential after warming. DESIGN: Prospective study. MATERIALS AND METHODS: The preliminary study was performed, using zygotes previously vitrified at pronuclear stage (n¼78) and day 3 (n¼36). They were warmed once, randomly allocated to either of the two vitrification methods, Cryotop or Rapid-i and then revitrified. Postwarming survival rate and subsequent development at day 5 and 6 were compared between the two vitrification methods. The present study was clinically performed, using 962 blastocysts. They were randomly allocated either Cryotop (n¼768) or Rapid-i (n¼194) and vitrified-warmed. Survival rates and pregnancy rates of single blastocyst transfer were compared. In evaluation of pregnancy rate, over 39-year old patients were excluded. RESULTS: The preliminary study showed all 2PNs and day 3 embryos survived after warming in both methods. Blastocyst rate from 2PN after warming was not significantly different between Cryotop and Rapid-i (day 5: 6/35, 17.1% vs. 6/43, 14.0%, day 6: 9/35, 25.7% vs. 11/43, 25.6%, respectively). The evaluation of development to blastocyst from
Vol. 102, No. 3, Supplement, September 2014
day 3 embryos after warming also showed similar results between Cryotop and Rapid-i (day 5: 5/18, 27.0% vs. 9/18, 50.0%, day 6: 10/18, 55.6% vs. 12/18, 66.7%, respectively). In the present study, there were no differences in survival rates (752/768, 97.9% vs. 190/194, 97.9%) and pregnancy rates (268/488, 54.9% vs. 57/109, 52.3 %) between Cryotop and Rapid-i. CONCLUSION: The present study demonstrates that a newly developed device, Rapid-i, dose not impair not only developmental potential of 2PN, day 3 embryo and blastocyst after warming, but also subsequent pregnancy rate compared to a conventional Cryotop method. Therefore, it is concluded that Rapid-i which does not expose human embryos directly to liquid nitrogen is a favorable device for storage without the risk of crosscontamination.
pregnancy rate/thaw by number of oocytes thawed showed no significant differences (44.6% of 101 thaws of 5 vs 41.5% of176 thaws of 7 oocytes). CONCLUSION: This study establishes the efficiency and viability of cryopreserved oocytes in IVF cycles. Low oocyte numbers per lot did not negatively impact overall outcomes therefore vitrified donor oocytes add ease of cycle timing and eliminate cancellations due to poor donor response. Supported by: This work was supported, in part, by the Program in Reproductive and Adult Endocrinology, NICHD, NIH, Bethesda, MD.
O-358 Wednesday, October 22, 2014 04:15 PM
PRE-IMPLANTATION GENETIC SCREENED (PGS)ABNORMAL BLASTOCYSTS GIVE SIGNIFICANT LESS SURVIVAL AFTER VITRIFICATION AND WARMING. R. Dunn, C. Vanijgul, R. Mangal, S. Chauhan, L. Schenk, W.-S. Wun, G. Grunert. Fertility Specialists of Houston, Houston, TX.
BANKED FROZEN DONOR OOCYTES DEMONSTRATE HIGH EFFICIENCY IN CONVERSION TO LIVE BORN INFANTS; A COLLABORATIVE MULTI-SITE EXPERIENCE. K. Fru,a J. Cox,a J. Lim,b B. M. Berger,c H. Hayes,c J. H. Segars,a D. I. Hoffman.c aNICHD, NIH, Bethesda, MD; bShady Grove Fertility, Rockville, MD; cDEBUSA, Rockville, MD. OBJECTIVE: Previous studies have shown a <7% oocyte to live born infant rate in fresh donor cycles. The purpose of this study was to evaluate the efficiency of cryopreserved donor oocytes compared to those from fresh donor cycles. DESIGN: Retrospective cohort study. MATERIALS AND METHODS: Outcomes of all IVF cycles using previously cryopreserved donor oocytes from a single donor oocyte bank and performed at 19 partner ART institutions(1/2012-12/2013) were reviewed. These were compared to fresh donor IVF cycles performed at a large participant ART institution in 2012. Outcomes included cycle cancellation, number of oocytes thawed, number of cryopreserved supernumerary embryos, clinical pregnancy (CP: intrauterine gestational sac), ongoing pregnancy rates (OPR) and number of infants born. RESULTS: 626 cycles using thawed oocytes were initiated with a 93% continuation rate; 41% of embryo transfer (ET) cycles had supernumerary embryos for cryopreservation (n¼238). Live birth rate/cycle was 51% (fresh) vs 44% (thaw)yielding 456 vs 332 infants. The twin rate was 14% vs 28% for fresh vs thaw donor cycles. Given an expected vitrified embryo to infant conversion rate of 35%, an additional 516 vs 150 infants are expected from these fresh and thaw cycles respectively. Final #infants/# thawed M2 oocytes was therefore (332+150)/3881 or 12.4% compared to (456+516)/8638 or 11.7% in fresh donor oocyte cycles. As frozen oocytes were stored in lots of 5-9, comparisons of ongoing
Single Site Fresh Cycles 2012 # Initiated Cycles #M2 Oocytes #M2 Oocytes surviving thaw (%) # Cycles with ET (%) # Cancelled Cycles (%) CP/Cycle Initiated CP/ET Cycle OPR/Initiated cycle OPR/Transfer Cycle # Liveborn Infants Sets of Twins (%) # Cycles with Vitrified Blasts (%) # Vitrified Blasts Expected FET Delivery Rate Expected # FET infants #Infants/M2 Oocytes
FERTILITY & STERILITYÒ
1088 8638 899 (83%) 132 (12%) 51% 61% 42% 51% 456 60 (14%) 538 (60%) 1476 35% 516 11.70%
Multi-site Cryo-Bank Cycles (2012-2013) 626 3881 3337 (86%) 582 (93%) 44 (7%) 46% 50% 41% 44% 332 71(28%) 238 (41%) 428 35% 150 12.40%
O-359 Wednesday, October 22, 2014 04:30 PM
OBJECTIVE: Aneuploid embryos are general considered not fruitful, i.e. arrested, degenerated. For embryos develop to blastocyst stage, these embryos have passed through maternal genomic activity to embryonic genomic activity and keep developing. Do these PGS abnormal blastocysts survive vitrification and warming as good as normal blastocysts do? In addition, general hypothesis is that developmental speed reflects the healthy status of embryos (1). Does day-5 biopsied blastocyst survive better than day-6 biopsied blastocyst does? This study examines the survival of vitrified blastocysts between PGS normal and abnormal embryos and the survival between day 5 and day 6 biopsied blastocysts. DESIGN: A retrospective study. MATERIALS AND METHODS: For PGS normal blastocysts, the frozen embryo transfer is usually 2 hours after warming. For PGS abnormal blastocysts, the examination of survival is 2 hours after warming then discarded. The definition of survival after2-hours warming is that at least half numbers of blastomeres are clear and shining. Those PGS cycles with frozen embryo transfer during period 1/1/2013 – 3/31/2014 are included. The age distribution is 22-45 with mean of 37.3. Totally 278 PGS normal embryos and 112 abnormal embryos from 416 cycles are included in the study. For trophectoderm biopsy, only those embryos develop to full or further advance blastocyst stage are biopsied on day 5. Other slow development blastocysts are biopsied on day 6. Fisher’s Exact Tests are used for analysis.
Biopsy date
PGS normal # survival/total (%)
PGS abnormal #survival/total (%)
Comparison between normal vs. abnormal
Day 5 Day 6 Analysis
147/148 (99%) 120/130 (92%) p <0.005
16/20 (80%) 66/92 (71%) n.s.
p <0.0001 p <0.0001 -
n.s.: not significant
RESULTS: The summary of results is in following table. CONCLUSION: 1. The survival of PGS abnormal blastocysts is significantly less than the PGS normal blastocysts does on both day 5 and day 6 biopsied blastocysts. 2. By comparison of PGS normal blastocysts, the survival of day 6 biopsied blastocysts is significantly less than day 6 biopsied blastocysts does. 3. By comparison of PGS abnormal blastocysts, there is no significant difference on the survival between day 5 and day6 biopsied blastocysts. It suggests PGS abnormality is more detrimental than slow blastocyst development. The results support that embryo growth rate affect the outcome of warmed blastocysts (Hashimoto et al, 2013). It also indicates there is a natural selection for normal and healthy blastocyst to avoid un-fruitful pregnancy.
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CRYOPRESERVATION AND FROZEN EMBRYO TRANSFER II O-356 Wednesday, October 22, 2014 03:45 PM EFFICACY AND EFFICIENCY OF THE ‘‘UNIVERSAL WARMING PROTOCOL’’: MULTICENTER RANDOMIZED CONTROLLED STUDY ON HUMAN SLOW FROZEN OOCYTES. L. Parmegiani,a C. Garello,b A. Revelli,b L. Criscuoli,c S. Dabizzi,c R. Gualtieri,d R. Talevi,d M. Filicori.a aReproductive Medicine Unit, GynePro Medical Centers, Bologna, BO, Italy; bLivet Clinic, Turin, TO, Italy; cCentre of Reproductive Medicine,, Careggi University Hospital, Florence, FI, Italy; d Dipartimento di Biologia, Universita di Napoli ‘‘Federico II’’, Naples, Italy. OBJECTIVE: Human reproductive cells are cryopreserved via slow freezing (SF) or vitrification (VIT). In a previously published pilot-
e122
ASRM Abstracts
study, it has already been demonstrated that it is possible to warm SF oocytes by using warming solution for VIT and also to increase their survival rates (1). The aim of the present study was to confirm the efficacy of the ‘‘rapid warming protocol’’ for SF oocytes in a multicenter trial and to compare its efficiency with the standard rapid thawing protocols. DESIGN: Three Assisted Reproductive (AR) centers in Italy were involved in this study. This was a prospective study on 414 oocytes randomized for conventional rapid thawing (RT) via two ready-to-use thawing media (Oocyte Thawing-Kit Origio–group A; CSC Thawing Medium Kit – group B), or rapid warming (RW) via VIT warming solutions (Vitrification Warming-Kit Sage – group C). MATERIALS AND METHODS: Primary endpoint was the morphological assessment of survival at 2 hours. Some of the surviving oocytes were divided into 3 sub-groups: i) parthenogenetically activated and observed by time-lapse microscopy ii) fixed and observed by confocal microscopy, iii) evaluated by polarized light for the presence of the meiotic spindle. Secondary endpoints were parthenogenetic activation and development, the assessment of the markers of oocyte preservation by confocal microscopy and the spindle presence by polarized light. RESULTS: Survival rate was 80.7% (71/88) in group A, 60.4% (125/ 207) in group B and 91.6 (109/119) in group C. Survival rate was significantly higher in group C (P¼0.036 vs group A and P<0.001 vs group B), in which the SF oocytes were warmed via VIT warming solutions. Survival rate was also significantly higher in group A vs group B (P¼0.001) Parthenogenetic activation and meiotic spindle presence were not significantly different in the three groups. At the time of writing this abstract the evaluation by confocal microscopy is still in progress. CONCLUSION: The findings of this study seem to confirm that is possible to optimize costs and increase the survival rate by using the same warming protocol for both SF and VIT oocytes. This ‘‘universal warming protocol’’ is potentially applicable to all kind of slow frozen reproductive cells, such as zygotes, cleavage stage embryos and blastocysts. Supported by: Thawing and warming solution were kindly supplied by Origio. O-357 Wednesday, October 22, 2014 04:00 PM COMPARISON OF VITRIFICATION DEVICES FOR HUMAN EMBRYOS BETWEEN OPEN AND CLOSED SYSTEM. S. Mizuno,a A. Ohgaki,a H. Matsumoto,a A. Fukuda,a Y. Morimoto.b aIVF Osaka Clinic, Higashi-Osaka, Osaka, Japan; bIVF Namba Clinic, Nishi-ku Osaka, Osaka, Japan. OBJECTIVE: Vitrification is a freezing method that is applied in almost every IVF clinic in Japan. In our clinic, human embryos are vitrified by Cryotop method in which the solution containing embryos is directly exposed to liquid nitrogen. This method does not completely eliminate the risk of cross-contamination during their storage. Therefore, the closed vitrification device, Rapid-i, has been developed to solve this problem. In the present study, survival rate and subsequent development of warmed embryos were compared between Cryotop and Rapid-i, to evaluate if embryos can be safely cryopreserved by Rapid-i without sacrificing their potential after warming. DESIGN: Prospective study. MATERIALS AND METHODS: The preliminary study was performed, using zygotes previously vitrified at pronuclear stage (n¼78) and day 3 (n¼36). They were warmed once, randomly allocated to either of the two vitrification methods, Cryotop or Rapid-i and then revitrified. Postwarming survival rate and subsequent development at day 5 and 6 were compared between the two vitrification methods. The present study was clinically performed, using 962 blastocysts. They were randomly allocated either Cryotop (n¼768) or Rapid-i (n¼194) and vitrified-warmed. Survival rates and pregnancy rates of single blastocyst transfer were compared. In evaluation of pregnancy rate, over 39-year old patients were excluded. RESULTS: The preliminary study showed all 2PNs and day 3 embryos survived after warming in both methods. Blastocyst rate from 2PN after warming was not significantly different between Cryotop and Rapid-i (day 5: 6/35, 17.1% vs. 6/43, 14.0%, day 6: 9/35, 25.7% vs. 11/43, 25.6%, respectively). The evaluation of development to blastocyst from
Vol. 102, No. 3, Supplement, September 2014
day 3 embryos after warming also showed similar results between Cryotop and Rapid-i (day 5: 5/18, 27.0% vs. 9/18, 50.0%, day 6: 10/18, 55.6% vs. 12/18, 66.7%, respectively). In the present study, there were no differences in survival rates (752/768, 97.9% vs. 190/194, 97.9%) and pregnancy rates (268/488, 54.9% vs. 57/109, 52.3 %) between Cryotop and Rapid-i. CONCLUSION: The present study demonstrates that a newly developed device, Rapid-i, dose not impair not only developmental potential of 2PN, day 3 embryo and blastocyst after warming, but also subsequent pregnancy rate compared to a conventional Cryotop method. Therefore, it is concluded that Rapid-i which does not expose human embryos directly to liquid nitrogen is a favorable device for storage without the risk of crosscontamination.
pregnancy rate/thaw by number of oocytes thawed showed no significant differences (44.6% of 101 thaws of 5 vs 41.5% of176 thaws of 7 oocytes). CONCLUSION: This study establishes the efficiency and viability of cryopreserved oocytes in IVF cycles. Low oocyte numbers per lot did not negatively impact overall outcomes therefore vitrified donor oocytes add ease of cycle timing and eliminate cancellations due to poor donor response. Supported by: This work was supported, in part, by the Program in Reproductive and Adult Endocrinology, NICHD, NIH, Bethesda, MD.
O-358 Wednesday, October 22, 2014 04:15 PM
PRE-IMPLANTATION GENETIC SCREENED (PGS)ABNORMAL BLASTOCYSTS GIVE SIGNIFICANT LESS SURVIVAL AFTER VITRIFICATION AND WARMING. R. Dunn, C. Vanijgul, R. Mangal, S. Chauhan, L. Schenk, W.-S. Wun, G. Grunert. Fertility Specialists of Houston, Houston, TX.
BANKED FROZEN DONOR OOCYTES DEMONSTRATE HIGH EFFICIENCY IN CONVERSION TO LIVE BORN INFANTS; A COLLABORATIVE MULTI-SITE EXPERIENCE. K. Fru,a J. Cox,a J. Lim,b B. M. Berger,c H. Hayes,c J. H. Segars,a D. I. Hoffman.c aNICHD, NIH, Bethesda, MD; bShady Grove Fertility, Rockville, MD; cDEBUSA, Rockville, MD. OBJECTIVE: Previous studies have shown a <7% oocyte to live born infant rate in fresh donor cycles. The purpose of this study was to evaluate the efficiency of cryopreserved donor oocytes compared to those from fresh donor cycles. DESIGN: Retrospective cohort study. MATERIALS AND METHODS: Outcomes of all IVF cycles using previously cryopreserved donor oocytes from a single donor oocyte bank and performed at 19 partner ART institutions(1/2012-12/2013) were reviewed. These were compared to fresh donor IVF cycles performed at a large participant ART institution in 2012. Outcomes included cycle cancellation, number of oocytes thawed, number of cryopreserved supernumerary embryos, clinical pregnancy (CP: intrauterine gestational sac), ongoing pregnancy rates (OPR) and number of infants born. RESULTS: 626 cycles using thawed oocytes were initiated with a 93% continuation rate; 41% of embryo transfer (ET) cycles had supernumerary embryos for cryopreservation (n¼238). Live birth rate/cycle was 51% (fresh) vs 44% (thaw)yielding 456 vs 332 infants. The twin rate was 14% vs 28% for fresh vs thaw donor cycles. Given an expected vitrified embryo to infant conversion rate of 35%, an additional 516 vs 150 infants are expected from these fresh and thaw cycles respectively. Final #infants/# thawed M2 oocytes was therefore (332+150)/3881 or 12.4% compared to (456+516)/8638 or 11.7% in fresh donor oocyte cycles. As frozen oocytes were stored in lots of 5-9, comparisons of ongoing
Single Site Fresh Cycles 2012 # Initiated Cycles #M2 Oocytes #M2 Oocytes surviving thaw (%) # Cycles with ET (%) # Cancelled Cycles (%) CP/Cycle Initiated CP/ET Cycle OPR/Initiated cycle OPR/Transfer Cycle # Liveborn Infants Sets of Twins (%) # Cycles with Vitrified Blasts (%) # Vitrified Blasts Expected FET Delivery Rate Expected # FET infants #Infants/M2 Oocytes
FERTILITY & STERILITYÒ
1088 8638 899 (83%) 132 (12%) 51% 61% 42% 51% 456 60 (14%) 538 (60%) 1476 35% 516 11.70%
Multi-site Cryo-Bank Cycles (2012-2013) 626 3881 3337 (86%) 582 (93%) 44 (7%) 46% 50% 41% 44% 332 71(28%) 238 (41%) 428 35% 150 12.40%
O-359 Wednesday, October 22, 2014 04:30 PM
OBJECTIVE: Aneuploid embryos are general considered not fruitful, i.e. arrested, degenerated. For embryos develop to blastocyst stage, these embryos have passed through maternal genomic activity to embryonic genomic activity and keep developing. Do these PGS abnormal blastocysts survive vitrification and warming as good as normal blastocysts do? In addition, general hypothesis is that developmental speed reflects the healthy status of embryos (1). Does day-5 biopsied blastocyst survive better than day-6 biopsied blastocyst does? This study examines the survival of vitrified blastocysts between PGS normal and abnormal embryos and the survival between day 5 and day 6 biopsied blastocysts. DESIGN: A retrospective study. MATERIALS AND METHODS: For PGS normal blastocysts, the frozen embryo transfer is usually 2 hours after warming. For PGS abnormal blastocysts, the examination of survival is 2 hours after warming then discarded. The definition of survival after2-hours warming is that at least half numbers of blastomeres are clear and shining. Those PGS cycles with frozen embryo transfer during period 1/1/2013 – 3/31/2014 are included. The age distribution is 22-45 with mean of 37.3. Totally 278 PGS normal embryos and 112 abnormal embryos from 416 cycles are included in the study. For trophectoderm biopsy, only those embryos develop to full or further advance blastocyst stage are biopsied on day 5. Other slow development blastocysts are biopsied on day 6. Fisher’s Exact Tests are used for analysis.
Biopsy date
PGS normal # survival/total (%)
PGS abnormal #survival/total (%)
Comparison between normal vs. abnormal
Day 5 Day 6 Analysis
147/148 (99%) 120/130 (92%) p <0.005
16/20 (80%) 66/92 (71%) n.s.
p <0.0001 p <0.0001 -
n.s.: not significant
RESULTS: The summary of results is in following table. CONCLUSION: 1. The survival of PGS abnormal blastocysts is significantly less than the PGS normal blastocysts does on both day 5 and day 6 biopsied blastocysts. 2. By comparison of PGS normal blastocysts, the survival of day 6 biopsied blastocysts is significantly less than day 6 biopsied blastocysts does. 3. By comparison of PGS abnormal blastocysts, there is no significant difference on the survival between day 5 and day6 biopsied blastocysts. It suggests PGS abnormality is more detrimental than slow blastocyst development. The results support that embryo growth rate affect the outcome of warmed blastocysts (Hashimoto et al, 2013). It also indicates there is a natural selection for normal and healthy blastocyst to avoid un-fruitful pregnancy.
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