- Email: [email protected]
Contractions during the expanded blastocyst stage decrease the success rate of frozen-thawed blastocyst transfer: time-lapse video analysis
currently there is limited data to support how long an embryo should stay at certain developmental stages before engaging in cytokinesis relative to embryo quality. Wong et. al described cytokinesis intervals of day 2 embryos to predict blastocyst formation. The focus of this study is to determine key indicators of embryo development which indicate a successful clinical outcome. The EmbryoscopeÒ Fertilitech allows IVF labs to leave embryos undisturbed in culture while scoring, which it theorized to benefit embryo development. Dynamics of embryo cleavage and development are paramount to patient success. DESIGN: Retrospective analyses of patients (n¼69) having IVF and cultured in the EmbryoscopeÒ Fertilitech at the Fertility Centers of New England from April 2012 through October 2012. Embryos with 100 % implantation rate (IR) vs. embryos with 0 % IR were included in this study. MATERIALS AND METHODS: 69 patients: 57 had 0 % IR and 12 had 100 %. Patients with 100 % IR had an mean age of 31years while patients with 0% had a mean of 37 years. Average BMI for both groups is 24. RESULTS: Looking at rates of cleavage as it related to implantation we identified three key points of development: 3 cell embryos with an increased lag to the 4 cell stage similar to Wong et.al., cleavage from 4 to 5 cell showed a difference, but less important. The time lag between the morula and blastocyst stage seems to be an important indicator of implantation potential. Due to the small N, additional data collection is warranted. CONCLUSION: Further data is currently being collected to analyze and validate these findings. If an embryo has already cleaved by the recomended time scoring, the embryoscope allows us to determine the exact time of cytokinesis.
EMBRYO CULTURE P-339 Tuesday, October 15, 2013 IMPLANTATION, PREGNANCY AND SPONTANEOUS ABORTION RATES FOLLOWING TRANSFER OF FROZENTHAWED BLASTOCYSTS CULTURED IN SEQUENTIAL OR CONTINUOUS EMBRYO CULTURE MEDIA PRIOR TO CRYOPRESERVATION. M. A. Stout,a J. H. Lim,a T. S. Han,a M. J. Levy,a J. R. Graham,a M. J. Tucker.a,b aEmbryology, Shady Grove Fertility Reproductive Science Center, Rockville, MD; bEmbryology, Georgia Reproductive Specialists, Atlanta, GA. OBJECTIVE: Embryos cultured in continuous culture medium have been reported to produce a higher rate of day-5 blastocyst compared with embryos cultured in sequential media. The purpose of this study was to evaluate clinical outcomes (implantation, pregnancy and spontaneous abortion rates) following transfer of frozen-thawed blastocysts cultured in sequential or continuous culture medium. DESIGN: Retrospective analysis. MATERIALS AND METHODS: Frozen-thawed embryo transfer (FET) categorized as autologous, 1st FET and day-5 transfer performed at a single fertility practice were reviewed. Blastocysts cryopreserved fom January 2010 to June 2011 were cultured in a commercially available sequentialstep embryo culture media (SSC); then from September 2011 to December 2012 were cultured in continuous single-step embryo culture medium (CSC). Embryos cultured in SSC were removed from cleavage medium and transferred to blastocyst medium on day-4 of culture. Embryos cultured in CSC were uninterrupted for the duration of their culture. FET’s were divided into two groups by age (%35 and >35). Implantation rates were compared by Mann-Whitney rank sum test. Pregnancy and spontaneous abortion rates were compared by chi-square. RESULTS: Clinical outcomes were similar following transfer of frozenthawed blastocysts cultured in SSC or CSC embryo culture media (Table 1., P> 0.05).
CONCLUSION: Frozen-thawed blastocysts previously cultured in continuous culture medium resulted in similar clinical outcomes compared with blastocysts cultured in sequential culture media. However, with an increase in production of day-5 blastocysts following culture in continuous culture medium and positive trend seen here, this may ultimately result in higher cumulative pregnancy rates. P-340 Tuesday, October 15, 2013 CONTRACTIONS DURING THE EXPANDED BLASTOCYST STAGE DECREASE THE SUCCESS RATE OF FROZEN-THAWED BLASTOCYST TRANSFER: TIME-LAPSE VIDEO ANALYSIS. S. Watanabe,a M. Kamihata,a R. Matsunaga,a A. Kuwahata,a,b M. Ochi,a T. Horiuchi.c aOchi Yume Clinic Nagoya, Nagoya, Aichi, Japan; bKato Ladies Clinic, Shinjuku, Tokyo, Japan; cPrefectural University of Hiroshima Graduate School of Comprehensive Scientific Research, Shobara, Hiroshima, Japan. OBJECTIVE: Contractions are often observed during the expended blastocyst stage in blastocyst culture. Trophoblast destruction caused by such contractions may affect the blastocysts. This study evaluated the effect of blastocyst contraction number on frozen-thawed blastocyst transfer by analyzing the blastocyst development stage using an EmbryoScopeÔ(ES; Unisense FertiliTech) time-lapse incubator for embryos. DESIGN: Retrospective study. MATERIALS AND METHODS: Blastocyst culture were performed from October to December 2012. ES videos of 365 well-developed frozen blastocysts were analyzed, and the number of contractions per blastocyst between blastocele formation and freezing was recorded. ‘‘One contraction’’ was defined as a contraction that was observed prior to blastocele formation after the expanded blastocyst stage. When the lumen diameter was R160 mm and the inner cell mass lay within the blastocyst cavity, it was frozen as a good blastocyst. A frozen-thawed single blastocyst transfer was performed in the hormone replacement cycle to study the relationship between the number of contractions and the pregnancy rate. RESULTS: The number of blastocysts and the pregnancy rate in ‘‘0 contractions,’’ ‘‘1 contraction,’’ ‘‘2 contractions,’’ and ‘‘R3 contractions’’ groups was 111, 111, 76, and 48, respectively, and 68.8% (33/48), 49.1% (26/53), 40.0% (14/35), and 35.0% (7/20), respectively. Hence, pregnancy rate was significantly greater in the ‘‘0 contractions’’ group than in the other groups. CONCLUSION: The transfer success rate was excellent in blastocysts that had sufficient lumen diameter to be frozen and had no contractions during the expanded blastocyst stage. As the number of contractions increased, the pregnancy rate decreased. Blastocyst culture using ES needs to be observed in real time to enable the freezing of blastocysts when they reach the required size. Use of ES allows clinicians to minimize the effect of long-term cultivation on blastocysts and select high-quality embryos without the need for in vitro culture. P-341 Tuesday, October 15, 2013 EFFECT OF AN ABNORMAL FIRST CLEAVAGE ON EMBRYONIC DEVELOPMENT: TIME-LAPSE VIDEO ANALYSIS. S. Watanabe,a M. Kamihata,a R. Matsunaga,a A. Kuwahata,a,b M. Ochi,a T. Horiuchi.c a Ochi Yume Clinic Nagoya, Nagoya, Aichi, Japan; bKato Ladies Clinic, Shinjuku, Tokyo, Japan; cPrefectural University of Hiroshima Graduate School of Comprehensive Scientific Research, Shobara, Hiroshima, Japan. OBJECTIVE: The first cleavage is responsible for ovum quality. However, 3-cell embryos are occasionally observed on Day 2 of embryo culture. In videos using an EmbryoScope TM (ES; Unisense FertiliTech, Denmark) time-lapse incubator for embryos, we found that 3-cell embryos formed due to an abnormal first cleavage. We analyzed the videos of the first cleavage to study its incidence and the rate of good blastocysts obtained during subsequent culture.
TABLE 1.
Age
Culture
n¼
Implantation (SEM)
Pregnancy
n¼
Spontaneous Abortion
% 35 % 35 > 35 > 35
SSC CSC SSC CSC
332 275 226 137
43.3 % (2.9) 48.2 % (2.5) 35.1 % (2.8) 38.7 % (3.7)
50.3 % 55.3 % 46.0 % 48.2 %
172 164 108 79
13.4 % 11.0 % 22.2 % 12.7 %
FERTILITY & STERILITYÒ
S245
EMBRYO CULTURE P-339 Tuesday, October 15, 2013 IMPLANTATION, PREGNANCY AND SPONTANEOUS ABORTION RATES FOLLOWING TRANSFER OF FROZENTHAWED BLASTOCYSTS CULTURED IN SEQUENTIAL OR CONTINUOUS EMBRYO CULTURE MEDIA PRIOR TO CRYOPRESERVATION. M. A. Stout,a J. H. Lim,a T. S. Han,a M. J. Levy,a J. R. Graham,a M. J. Tucker.a,b aEmbryology, Shady Grove Fertility Reproductive Science Center, Rockville, MD; bEmbryology, Georgia Reproductive Specialists, Atlanta, GA. OBJECTIVE: Embryos cultured in continuous culture medium have been reported to produce a higher rate of day-5 blastocyst compared with embryos cultured in sequential media. The purpose of this study was to evaluate clinical outcomes (implantation, pregnancy and spontaneous abortion rates) following transfer of frozen-thawed blastocysts cultured in sequential or continuous culture medium. DESIGN: Retrospective analysis. MATERIALS AND METHODS: Frozen-thawed embryo transfer (FET) categorized as autologous, 1st FET and day-5 transfer performed at a single fertility practice were reviewed. Blastocysts cryopreserved fom January 2010 to June 2011 were cultured in a commercially available sequentialstep embryo culture media (SSC); then from September 2011 to December 2012 were cultured in continuous single-step embryo culture medium (CSC). Embryos cultured in SSC were removed from cleavage medium and transferred to blastocyst medium on day-4 of culture. Embryos cultured in CSC were uninterrupted for the duration of their culture. FET’s were divided into two groups by age (%35 and >35). Implantation rates were compared by Mann-Whitney rank sum test. Pregnancy and spontaneous abortion rates were compared by chi-square. RESULTS: Clinical outcomes were similar following transfer of frozenthawed blastocysts cultured in SSC or CSC embryo culture media (Table 1., P> 0.05).
CONCLUSION: Frozen-thawed blastocysts previously cultured in continuous culture medium resulted in similar clinical outcomes compared with blastocysts cultured in sequential culture media. However, with an increase in production of day-5 blastocysts following culture in continuous culture medium and positive trend seen here, this may ultimately result in higher cumulative pregnancy rates. P-340 Tuesday, October 15, 2013 CONTRACTIONS DURING THE EXPANDED BLASTOCYST STAGE DECREASE THE SUCCESS RATE OF FROZEN-THAWED BLASTOCYST TRANSFER: TIME-LAPSE VIDEO ANALYSIS. S. Watanabe,a M. Kamihata,a R. Matsunaga,a A. Kuwahata,a,b M. Ochi,a T. Horiuchi.c aOchi Yume Clinic Nagoya, Nagoya, Aichi, Japan; bKato Ladies Clinic, Shinjuku, Tokyo, Japan; cPrefectural University of Hiroshima Graduate School of Comprehensive Scientific Research, Shobara, Hiroshima, Japan. OBJECTIVE: Contractions are often observed during the expended blastocyst stage in blastocyst culture. Trophoblast destruction caused by such contractions may affect the blastocysts. This study evaluated the effect of blastocyst contraction number on frozen-thawed blastocyst transfer by analyzing the blastocyst development stage using an EmbryoScopeÔ(ES; Unisense FertiliTech) time-lapse incubator for embryos. DESIGN: Retrospective study. MATERIALS AND METHODS: Blastocyst culture were performed from October to December 2012. ES videos of 365 well-developed frozen blastocysts were analyzed, and the number of contractions per blastocyst between blastocele formation and freezing was recorded. ‘‘One contraction’’ was defined as a contraction that was observed prior to blastocele formation after the expanded blastocyst stage. When the lumen diameter was R160 mm and the inner cell mass lay within the blastocyst cavity, it was frozen as a good blastocyst. A frozen-thawed single blastocyst transfer was performed in the hormone replacement cycle to study the relationship between the number of contractions and the pregnancy rate. RESULTS: The number of blastocysts and the pregnancy rate in ‘‘0 contractions,’’ ‘‘1 contraction,’’ ‘‘2 contractions,’’ and ‘‘R3 contractions’’ groups was 111, 111, 76, and 48, respectively, and 68.8% (33/48), 49.1% (26/53), 40.0% (14/35), and 35.0% (7/20), respectively. Hence, pregnancy rate was significantly greater in the ‘‘0 contractions’’ group than in the other groups. CONCLUSION: The transfer success rate was excellent in blastocysts that had sufficient lumen diameter to be frozen and had no contractions during the expanded blastocyst stage. As the number of contractions increased, the pregnancy rate decreased. Blastocyst culture using ES needs to be observed in real time to enable the freezing of blastocysts when they reach the required size. Use of ES allows clinicians to minimize the effect of long-term cultivation on blastocysts and select high-quality embryos without the need for in vitro culture. P-341 Tuesday, October 15, 2013 EFFECT OF AN ABNORMAL FIRST CLEAVAGE ON EMBRYONIC DEVELOPMENT: TIME-LAPSE VIDEO ANALYSIS. S. Watanabe,a M. Kamihata,a R. Matsunaga,a A. Kuwahata,a,b M. Ochi,a T. Horiuchi.c a Ochi Yume Clinic Nagoya, Nagoya, Aichi, Japan; bKato Ladies Clinic, Shinjuku, Tokyo, Japan; cPrefectural University of Hiroshima Graduate School of Comprehensive Scientific Research, Shobara, Hiroshima, Japan. OBJECTIVE: The first cleavage is responsible for ovum quality. However, 3-cell embryos are occasionally observed on Day 2 of embryo culture. In videos using an EmbryoScope TM (ES; Unisense FertiliTech, Denmark) time-lapse incubator for embryos, we found that 3-cell embryos formed due to an abnormal first cleavage. We analyzed the videos of the first cleavage to study its incidence and the rate of good blastocysts obtained during subsequent culture.
TABLE 1.
Age
Culture
n¼
Implantation (SEM)
Pregnancy
n¼
Spontaneous Abortion
% 35 % 35 > 35 > 35
SSC CSC SSC CSC
332 275 226 137
43.3 % (2.9) 48.2 % (2.5) 35.1 % (2.8) 38.7 % (3.7)
50.3 % 55.3 % 46.0 % 48.2 %
172 164 108 79
13.4 % 11.0 % 22.2 % 12.7 %
FERTILITY & STERILITYÒ
S245