- Email: [email protected]
Effect of an abnormal first cleavage on embryonic development: 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
DESIGN: Retrospective study. MATERIALS AND METHODS: Of 1,344 normal fertilized embryos obtained from 612 cycles of 454 subjects who underwent ovum retrieval and blastocyst culture using the ES between July 2012 and February 2013, 139 with a 3-cell embryo on the morning of Day 2 were enrolled. Ovum retrieval was performed during the clomiphene or letrozole cycle for in vitro fertilization or intracytoplasmic sperm injection. Normal fertilized embryos were incubated in the ES. The video was taken once every 30 minutes. Time-lapse videos of the 3-cell embryos at the first cleavage on the morning of Day 2 (42 hours post-fertilization) were analyzed and divided into 2 groups, the normal group (2-cell embryos at the first cleavage found as 3-cell embryos among 4cell embryos) and the group of 3-cell embryos at the first cleavage. The blastocyst culture period was up to 7 days. When the lumen diameter was R160 mm and the inner cell mass lay within the blastocyst cavity, it was frozen as a good blastocyst. The good blastocyst rate was compared between the groups. RESULTS: The number of embryos and the good blastocyst rate in the normal and 3-cell embryo groups were 88 and 51.1% (45/88) and 51 and 9.8% (5/51), respectively, and the results of the 3-cell embryo group were significantly lower. CONCLUSION: Time-lapse video analysis showed that 36.7% of the 3cell embryos on Day 2 had an abnormal first cleavage. Additionally, their good blastocyst rate was quite low. These results suggest that 3-cell embryos at Day 2 should be excluded from the embryo transfer process. P-342 Tuesday, October 15, 2013 TRANSFER OF EMBRYOS FOLLOWING CULTURE IN CONTINUOUS SINGLE-STEP OR SEQUENTIAL-STEP EMBRYO CULTURE MEDIA. M. A. Stout,a V. A. Cholewczynski,a S. G. Shukry,a G. S. Karman,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: Evaluation of clinical outcomes (blastocyst conversion, implantation and pregnancy rates, sex ratios and birth weights) following culture / transfer of embryos cultured in commercially available continuous single-step (CSC) medium compared with sequential-step (SSC) embryo culture media. DESIGN: Retrospective analysis. MATERIALS AND METHODS: Clinical outcomes were compared following culture / transfer of embryos in SSC (June 2010 to July 2011) or CSC (October 2011 to December 2012). No other changes in laboratory or clinical protocols noted during study. Embryos cultured in SSC were transferred to blastocyst medium on day-4 of culture, CSC remained uninterrupted. Blastocyst conversion, pregnancy and implantation rates from patients aged %48 were analyzed by chi square and Mann-Whitney rank sum test, respectively. Sex ratio and birth weight of singleton neonates following transfer to patients aged %43 were analyzed by chi square and Mann-Whitney rank sum test, respectively. RESULTS: Embryos cultured in CSC (n¼2264) compared with SSC (n¼1683) were found to have increased blastocyst conversion rate (24.6% 0.5 vs. 20.9% 0.5, respectively; P<0.001) but were similar in implantation rates (38.9% 0.9 vs. 38.3% 1.1, respectively; P¼0.756) and pregnancy rates (47.5% vs. 47.4%, respectively; P¼0.964). Embryos cultured in SSC (n¼502) and CSC (n¼237) were also similar in sex ratio (% male¼50.6% vs. 50.2%, respectively; P¼0.985) and birth weight (3234.7g 25.8 vs. 3213.9g 44.0, respectively; P¼0.560). CONCLUSION: Culture of embryos in CSC has previously been reported to increase the number of day-5 embryos available for transfer or vitrification. During this study, we report an increase in blastocyst conversion following culture in CSC compared with SSC. However, following transfer, embryos cultured in CSC and SSC were similar in implantation, pregnancy rates, sex ratio and birth weight. This increase in blastocyst development may be due to a reduction in embryo stress during manipulation / culture.
P-343 Tuesday, October 15, 2013 THE EFFECT OF CULTURE MEDIA ON EMBRYO QUALITY AND THE INCIDENCE OF CRYOPRESERVATION. C. R. McCann, R. Halverson, J. Early, L. B. Davis, G. D. Ball. Seattle Reproductive Medicine, Seattle, WA. OBJECTIVE: Preliminary trials suggested that culture media had a notable effect on rate of embryo development, quality of embryos and number of high quality blastocysts available for cryopreservation. The purpose of
S246
ASRM Abstracts
this study was to compare embryo development in Sage versus Global culture media. DESIGN: Retrospective cohort study. Autologous patients (n¼336) under the age of 38 had a total of 2991 embryos cultured in Sage sequential step media. Autologous patients (n¼324) under the age of 38 had 2857 embryos cultured in Global Total single step media. All other culture conditions remained constant. Data was collected from July 2011 to January 2013. Only cycles resulting in an embryo transfer were included. MATERIALS AND METHODS: The groups were assessed for quality of embryos transferred (based on SART criteria), frequency of elective single embryo transfer (eSET), incidence of cryopreservation, clinical pregnancy rate, and multiple pregnancy rate. Standard culture conditions included 6% O2 and 6% CO2 at 37 C. RESULTS: The percentage of good quality embryos transferred significantly increased from 57% in Sage media to 75% in Global media (p¼0.001). The percentage of patients who had additional embryos to freeze significantly increased from 51% in Sage to 59% in Global (p¼0.03). The frequency of eSET increased from 31% in Sage to 37% in Global, although this difference was not significant (p>0.05). A small increase in clinical pregnancy rate and a slight decrease in multiple pregnancy rate were noted but these differences were not significant (p>0.05). CONCLUSION: Under the conditions chosen for culture in our laboratory, medium choice played a significant role in both embryo quality and incidence of cryopreservation. The availability of more high quality embryos is key in our effort to offer eSET to more patients while maintaining high success rates. A higher yield of embryos available for cryopreservation results in an overall better outcome for IVF patients. P-344 Tuesday, October 15, 2013 COMPARISON OF ANEUPLOIDY RATES BETWEEN DAY 5 AND DAY 6 HUMAN BLASTOCYSTS. F. Balmir,a M. Persad,a J. Jenkins,b J. R. Stelling.a aObstetrics and Gynecology, Stony Brook University Hospital, Stony Brook, NY; bReproductive Endocrinology, Reproductive Specialists of New York, Mineola, NY. OBJECTIVE: To compare rates of aneuploidy between human embryos biopsied on Day 5 or Day 6 using array comparative genomic hybridization (aCGH) to diagnose aneuploidy. DESIGN: This is a retrospective study in an in vitro fertilization laboratory. MATERIALS AND METHODS: A total of 245 embryos were included from 57 patients undergoing in vitro fertilization with embryo biopsies and analysis on Day 5 or Day 6 via aCGH between 01/01/2010 and 03/ 24/13. 245 embryos were identified and reviewed. Multiple blastomeres were biopsied on Day 5 or Day 6 of fertilization. Aneuploidy screening by aCGH for all 23 pairs of chromosomes was performed. A Chi square analysis was used to compare the rate of aneuploidy on Day 5 versus Day 6. RESULTS: Among the 245 embryos analyzed, 35.4% (85) were euploid and 56.7% (133) were aneuploid. The rate of aneuploidy on Day 5 was 59.39% compared to 66.33% seen on Day 6. The rate of euploidy on Day 5 was 40.6% versus that of 36.66% on Day 6. 8.33% of the total embryos could not be analyzed. CONCLUSION: There was no significant difference between the rate of aneuploidy on blastocysts biopsied on Day 5 versus Day 6, nor was there a significant difference between the rate of euploidy on Day 5 versus Day 6.
P-345 Tuesday, October 15, 2013 LOW LABORATORY RELATIVE HUMIDITIES APPEAR TO AFFECT INITIAL CONCEPTION RATES BUT NOT ON-GOING PREGNANCIES IN AN ART PROGRAM. S. Prien, R. Klukarni, L. Penrose. Ob/Gyn, Texas Tech University Health Sciences Center, Lubbock, TX. OBJECTIVE: It is also well established that well controlled humidity is an essential part of assisted reproductive technologies (ART) culture conditions and necessary for optimum embryo development. However, it is unclear to what extent lab relative humidity (RH), especially low RH, might impact the culture environment. For the last 24 months, environmental conditions in the Southwestern U.S. have produced extended periods of extremely low RH, which has resulted in lower overall building RH at this facility and providing an opportunity to examine the role of lab RH on culture conditions.
Vol. 100, No. 3, Supplement, September 2013
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
DESIGN: Retrospective study. MATERIALS AND METHODS: Of 1,344 normal fertilized embryos obtained from 612 cycles of 454 subjects who underwent ovum retrieval and blastocyst culture using the ES between July 2012 and February 2013, 139 with a 3-cell embryo on the morning of Day 2 were enrolled. Ovum retrieval was performed during the clomiphene or letrozole cycle for in vitro fertilization or intracytoplasmic sperm injection. Normal fertilized embryos were incubated in the ES. The video was taken once every 30 minutes. Time-lapse videos of the 3-cell embryos at the first cleavage on the morning of Day 2 (42 hours post-fertilization) were analyzed and divided into 2 groups, the normal group (2-cell embryos at the first cleavage found as 3-cell embryos among 4cell embryos) and the group of 3-cell embryos at the first cleavage. The blastocyst culture period was up to 7 days. When the lumen diameter was R160 mm and the inner cell mass lay within the blastocyst cavity, it was frozen as a good blastocyst. The good blastocyst rate was compared between the groups. RESULTS: The number of embryos and the good blastocyst rate in the normal and 3-cell embryo groups were 88 and 51.1% (45/88) and 51 and 9.8% (5/51), respectively, and the results of the 3-cell embryo group were significantly lower. CONCLUSION: Time-lapse video analysis showed that 36.7% of the 3cell embryos on Day 2 had an abnormal first cleavage. Additionally, their good blastocyst rate was quite low. These results suggest that 3-cell embryos at Day 2 should be excluded from the embryo transfer process. P-342 Tuesday, October 15, 2013 TRANSFER OF EMBRYOS FOLLOWING CULTURE IN CONTINUOUS SINGLE-STEP OR SEQUENTIAL-STEP EMBRYO CULTURE MEDIA. M. A. Stout,a V. A. Cholewczynski,a S. G. Shukry,a G. S. Karman,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: Evaluation of clinical outcomes (blastocyst conversion, implantation and pregnancy rates, sex ratios and birth weights) following culture / transfer of embryos cultured in commercially available continuous single-step (CSC) medium compared with sequential-step (SSC) embryo culture media. DESIGN: Retrospective analysis. MATERIALS AND METHODS: Clinical outcomes were compared following culture / transfer of embryos in SSC (June 2010 to July 2011) or CSC (October 2011 to December 2012). No other changes in laboratory or clinical protocols noted during study. Embryos cultured in SSC were transferred to blastocyst medium on day-4 of culture, CSC remained uninterrupted. Blastocyst conversion, pregnancy and implantation rates from patients aged %48 were analyzed by chi square and Mann-Whitney rank sum test, respectively. Sex ratio and birth weight of singleton neonates following transfer to patients aged %43 were analyzed by chi square and Mann-Whitney rank sum test, respectively. RESULTS: Embryos cultured in CSC (n¼2264) compared with SSC (n¼1683) were found to have increased blastocyst conversion rate (24.6% 0.5 vs. 20.9% 0.5, respectively; P<0.001) but were similar in implantation rates (38.9% 0.9 vs. 38.3% 1.1, respectively; P¼0.756) and pregnancy rates (47.5% vs. 47.4%, respectively; P¼0.964). Embryos cultured in SSC (n¼502) and CSC (n¼237) were also similar in sex ratio (% male¼50.6% vs. 50.2%, respectively; P¼0.985) and birth weight (3234.7g 25.8 vs. 3213.9g 44.0, respectively; P¼0.560). CONCLUSION: Culture of embryos in CSC has previously been reported to increase the number of day-5 embryos available for transfer or vitrification. During this study, we report an increase in blastocyst conversion following culture in CSC compared with SSC. However, following transfer, embryos cultured in CSC and SSC were similar in implantation, pregnancy rates, sex ratio and birth weight. This increase in blastocyst development may be due to a reduction in embryo stress during manipulation / culture.
P-343 Tuesday, October 15, 2013 THE EFFECT OF CULTURE MEDIA ON EMBRYO QUALITY AND THE INCIDENCE OF CRYOPRESERVATION. C. R. McCann, R. Halverson, J. Early, L. B. Davis, G. D. Ball. Seattle Reproductive Medicine, Seattle, WA. OBJECTIVE: Preliminary trials suggested that culture media had a notable effect on rate of embryo development, quality of embryos and number of high quality blastocysts available for cryopreservation. The purpose of
S246
ASRM Abstracts
this study was to compare embryo development in Sage versus Global culture media. DESIGN: Retrospective cohort study. Autologous patients (n¼336) under the age of 38 had a total of 2991 embryos cultured in Sage sequential step media. Autologous patients (n¼324) under the age of 38 had 2857 embryos cultured in Global Total single step media. All other culture conditions remained constant. Data was collected from July 2011 to January 2013. Only cycles resulting in an embryo transfer were included. MATERIALS AND METHODS: The groups were assessed for quality of embryos transferred (based on SART criteria), frequency of elective single embryo transfer (eSET), incidence of cryopreservation, clinical pregnancy rate, and multiple pregnancy rate. Standard culture conditions included 6% O2 and 6% CO2 at 37 C. RESULTS: The percentage of good quality embryos transferred significantly increased from 57% in Sage media to 75% in Global media (p¼0.001). The percentage of patients who had additional embryos to freeze significantly increased from 51% in Sage to 59% in Global (p¼0.03). The frequency of eSET increased from 31% in Sage to 37% in Global, although this difference was not significant (p>0.05). A small increase in clinical pregnancy rate and a slight decrease in multiple pregnancy rate were noted but these differences were not significant (p>0.05). CONCLUSION: Under the conditions chosen for culture in our laboratory, medium choice played a significant role in both embryo quality and incidence of cryopreservation. The availability of more high quality embryos is key in our effort to offer eSET to more patients while maintaining high success rates. A higher yield of embryos available for cryopreservation results in an overall better outcome for IVF patients. P-344 Tuesday, October 15, 2013 COMPARISON OF ANEUPLOIDY RATES BETWEEN DAY 5 AND DAY 6 HUMAN BLASTOCYSTS. F. Balmir,a M. Persad,a J. Jenkins,b J. R. Stelling.a aObstetrics and Gynecology, Stony Brook University Hospital, Stony Brook, NY; bReproductive Endocrinology, Reproductive Specialists of New York, Mineola, NY. OBJECTIVE: To compare rates of aneuploidy between human embryos biopsied on Day 5 or Day 6 using array comparative genomic hybridization (aCGH) to diagnose aneuploidy. DESIGN: This is a retrospective study in an in vitro fertilization laboratory. MATERIALS AND METHODS: A total of 245 embryos were included from 57 patients undergoing in vitro fertilization with embryo biopsies and analysis on Day 5 or Day 6 via aCGH between 01/01/2010 and 03/ 24/13. 245 embryos were identified and reviewed. Multiple blastomeres were biopsied on Day 5 or Day 6 of fertilization. Aneuploidy screening by aCGH for all 23 pairs of chromosomes was performed. A Chi square analysis was used to compare the rate of aneuploidy on Day 5 versus Day 6. RESULTS: Among the 245 embryos analyzed, 35.4% (85) were euploid and 56.7% (133) were aneuploid. The rate of aneuploidy on Day 5 was 59.39% compared to 66.33% seen on Day 6. The rate of euploidy on Day 5 was 40.6% versus that of 36.66% on Day 6. 8.33% of the total embryos could not be analyzed. CONCLUSION: There was no significant difference between the rate of aneuploidy on blastocysts biopsied on Day 5 versus Day 6, nor was there a significant difference between the rate of euploidy on Day 5 versus Day 6.
P-345 Tuesday, October 15, 2013 LOW LABORATORY RELATIVE HUMIDITIES APPEAR TO AFFECT INITIAL CONCEPTION RATES BUT NOT ON-GOING PREGNANCIES IN AN ART PROGRAM. S. Prien, R. Klukarni, L. Penrose. Ob/Gyn, Texas Tech University Health Sciences Center, Lubbock, TX. OBJECTIVE: It is also well established that well controlled humidity is an essential part of assisted reproductive technologies (ART) culture conditions and necessary for optimum embryo development. However, it is unclear to what extent lab relative humidity (RH), especially low RH, might impact the culture environment. For the last 24 months, environmental conditions in the Southwestern U.S. have produced extended periods of extremely low RH, which has resulted in lower overall building RH at this facility and providing an opportunity to examine the role of lab RH on culture conditions.
Vol. 100, No. 3, Supplement, September 2013