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Trophectoderm grade is a better predictor than inner cell mass grade for selecting blastocysts for embryo transfer
EMBRYO CULTURE
P-185 Tuesday, October 23, 2012 TROPHECTODERM GRADE IS A BETTER PREDICTOR THAN INNER CELL MASS GRADE FOR SELECTING BLASTOCYSTS N. Fukunaga,a,b,c FOR EMBRYO TRANSFER. H. Ohno,a,b R. Nagai,a,b H. Kitasaka,a,b M. Kato,a,b Y. Asada.a,b,c aAsada Ladies Nagoya Clinic, Nagoya, Aichi, Japan; bAsada Ladies Kachigawa Clinic, Kasugai, Aichi, Japan; cThe Asada Institute for Reproductive Medicine, Kasugai, Aichi, Japan. OBJECTIVE: In cases where good-quality blastocysts with a Gardner score above 3BB are not obtained, we also freeze blastocysts of grade C. We analyzed whether blastocysts with an inner cell mass (ICM) and trophectoderm (TE) of grade C show a difference in the rates of pregnancy and abortion after embryo transfer compared with blastocysts having an ICM and TE of grades A or B. DESIGN: Retrospective study. MATERIALS AND METHODS: We conducted a retrospective analysis of all patients (n¼2993 transfers, 1589 patients) who had received a fresh or a frozen–thawed single blastocyst, transferred on day 5, at our clinic between January 2007 and March 2011. The blastocysts were graded according to the Gardner grading system in which three parameters are scored: the degree of blastocoele expansion and hatching status (1–6); size and compactness of the ICM (highest score A, then B, and C); and the cohesiveness and number of TE cells (A, B, and C). We graded the ICM and TE and compared the rates of pregnancy and abortion for blastocysts of grades A, B, and C. RESULTS: The rates of pregnancy per embryo transfer cycle for ICMs of grades A, B, and C were 45.1% (829/1838), 41.7% (454/1089), and 39.4% (26/66), and abortion rates were 19.8% (164/829), 24.5% (111/454), and 26.9% (7/26) respectively. The ICM grade did not have a statistically significant effect on the rate of pregnancy or abortion. In contrast, the rates of pregnancy per embryo transfer cycle for TE grades of A, B, and C were 48.8% (488/1000), 42.4% (786/1853), and 25.0% (35/140), respectively. For grade C TE, the rate of pregnancy was significantly lower than that for TEs of grade A or B (P<0.05). The rates of abortion for TEs of grades A, B, and C were 17.8% (87/488), 23.4% (184/786), and 31.4% (11/35), respectively. Embryos with a grade C TE showed a significantly higher rate of abortion compared with those with a grade A TE (P<0.05). CONCLUSION: We found that the grade of TE was a better predictor of implantation than ICM morphology.
P-186 Tuesday, October 23, 2012 SINGLE-STEP VS. TWO-STEP MEDIA: ARE THERE ANY DIFFERENCES IN OUTCOME AND IMPLANTATION? J. Liebermann, E. J. Pelts, R. Brohammer, Y. Wagner, C. Sipe, K. Lederer. Fertility Centers of Illinois, Chicago, IL. OBJECTIVE: The culture of embryos has been approached by two culture protocols. The two-step protocol is thought to mimic the physiological interaction between specific developmental stages of the embryo within the oviduct and uterus. The opposing system in based on the ‘let the embryo choose’ principal that is analogous to a single-step culture medium. By culturing sibling oocytes in both media under the same culture conditions, we compared both groups with respect to outcome and implantation. DESIGN: Retrospective analysis. MATERIALS AND METHODS: A single-step medium (GLOBAL, IVFonline, Guilford, CT) was compared to a two-step media from SAGE (Trumbull, CT). Embryo culture was performed as group culture in 4-well dishes under oil and 5% oxygen. On day-1, following 2pn-check, sibling zygote cohorts from 181 patients (average age of 33.54.5) were randomly allocated into the SAGE or GLOBAL culture media. 953 embryos were cultured in the SAGE, whereas 1049 embryos were cultured in GLOBAL. 58 transfers from the SAGE system were compared to 123 transfers from the GLOBAL system. Clinical pregnancies showing FCA activity as determined by ultrasound. Chi square test was used for statistical analysis. RESULTS: From both culture systems an average of 1.60.5 embryos were transferred. Almost 70% of the transfers (123/181) had embryos selected from the GLOBAL culture system. Overall, the total number of embryos cryopreserved and transferred was significantly higher in the GLOBAL group (57.3%; 601/1049; P<.04) compared to the SAGE group
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ASRM Abstracts
(47.6%; 454/953). Clinical pregnancy was slightly, but not significantly higher in SAGE (58.6%; 34/58; P<.92) than in GLOBAL (56.1%; 69/ 123). Furthermore, implantation was marginal, but not significant higher in SAGE (46.1%; 42/91; P<.75) compared to GLOBAL (42%; 82/196). CONCLUSION: This trial demonstrates that embryos selected from a single-step media provide the same potential in achieving good clinical pregnancy and implantation rates compared to their sibling embryos cultured in a two-step media.
P-187 Tuesday, October 23, 2012 EFFECT OF LIF ON HUMAN EMBRYOS. A. Zhioua,a N. Mami,a S. Fourati,a H. Elloumi,a M. Khrouf,b F. Zhioua.b aCenter of Reproductive Biology, Center of Reproductive Biology, Tunis, Tunisia; bDepartment of Gynecology Obstetrics, Department of Gynecology Obstetrics, Tunis, Tunisia. OBJECTIVE: Leukemia inhibitor factor (LIF) is a cytokine with a molecular weight of 38-67KDa, which is expressed at highest concentrations in both maternal endometrial glands and extra embryonic membranes of the embryo, at about the implantation stage. The purpose of this study is to assess whether addition of LIF into a specific culture media will improve the embryo quality, susceptible to enhance pregnancy rate following embryo transfer. DESIGN: This is a blind prospective study conducted in our IVF laboratory, during the months of March and April 2012, on 338 oocytes received the day of ovum pick-ups of 50 women underwent the ICSI program. MATERIALS AND METHODS: The woman age ranges from 21 to 41 years. The oocyte cohort of each woman is randomly divided into two groups, after the microinjection: Group 1: oocytes exposed to LIF, Group 2: oocytes not exposed to LIF. Recombinant human LIF lyophilized (INVITROGEN, GIBCO) is added to microdroplets of embryo culture media (G1 Plus, VITROLIFE) at a concentration of 2.5microg/ml. These are containing the microinjected oocytes placed in culture for 2 or 3 days. We compare the embryo quality, between the exposed and the non exposed embryos to LIF at day 2 and day 3 (blastomere number, regularity and fragmentation rate). RESULTS: Exposure to LIF during the early stages of embryo development shows a statistically significant improvement in the embryo quality especially at day 3, by increasing the number of embryos with regular blastomeres (80% (group1) vs 61% (group 2), P¼0.016) and lower fragmentation rate (83% (group1) vs 59% (group 2), P¼0.003). The number of eightcell embryo is also higher in the group of embryos exposed to LIF (27% vs 17%), but with no significant difference. CONCLUSION: These results indicate that LIF could have a benefic effect on embryo quality, especially at day 3, which may increase the chance of pregnancy after ICSI. This preliminary study deserves, however, to be complemented by a larger sample in order to confirm the results obtained.
P-188 Tuesday, October 23, 2012 AN OOCYTE SPECIFIC MEDIUM FOR THE FIRST 24 HOURS OF CULTURE IMPROVES DEVELOPMENT OF GOOD QUALITY J. Stevens,b A. Janesch,b BLASTOCYSTS. R. L. Krisher,a S. D. Degelos,c G. Koulianos,c W. B. Schoolcraft.b aNational Foundation for Fertility Research, Lone Tree, CO; bColorado Center for Reproductive Medicine, Lone Tree, CO; cCenter for Reproductive Medicine, Mobile, AL. OBJECTIVE: Although oocytes have unique requirements, typically embryo culture media are used for oocyte collection and initial culture in IVF cycles. The objective of this study was to evaluate the efficacy of a medium designed specifically for the needs of oocytes, and used for the initial 24 hours of culture to support subsequent blastocyst development. DESIGN: Research study. MATERIALS AND METHODS: Oocytes (n¼575) from each patient (n¼30) were randomly allocated at retrieval to Quinns Advantage Cleavage Medium (CM, SAGE) or a new, oocyte specific medium (Oocyte Handling Medium, OHM) for the initial 24 hours of culture. ICSI was performed in OHM or CM. Fertilized embryos from both groups were then transferred for the remainder of culture to commercial sequential media (SAGE). Embryos were cultured to D5 or D6, and blastocyst development assessed. Good quality embryos were either transferred fresh (D5) or frozen (D5 or D6). Twenty four patients have undergone blastocyst transfer (19 fresh, 5 frozen).
Vol. 98, No. 3, Supplement, September 2012
P-185 Tuesday, October 23, 2012 TROPHECTODERM GRADE IS A BETTER PREDICTOR THAN INNER CELL MASS GRADE FOR SELECTING BLASTOCYSTS N. Fukunaga,a,b,c FOR EMBRYO TRANSFER. H. Ohno,a,b R. Nagai,a,b H. Kitasaka,a,b M. Kato,a,b Y. Asada.a,b,c aAsada Ladies Nagoya Clinic, Nagoya, Aichi, Japan; bAsada Ladies Kachigawa Clinic, Kasugai, Aichi, Japan; cThe Asada Institute for Reproductive Medicine, Kasugai, Aichi, Japan. OBJECTIVE: In cases where good-quality blastocysts with a Gardner score above 3BB are not obtained, we also freeze blastocysts of grade C. We analyzed whether blastocysts with an inner cell mass (ICM) and trophectoderm (TE) of grade C show a difference in the rates of pregnancy and abortion after embryo transfer compared with blastocysts having an ICM and TE of grades A or B. DESIGN: Retrospective study. MATERIALS AND METHODS: We conducted a retrospective analysis of all patients (n¼2993 transfers, 1589 patients) who had received a fresh or a frozen–thawed single blastocyst, transferred on day 5, at our clinic between January 2007 and March 2011. The blastocysts were graded according to the Gardner grading system in which three parameters are scored: the degree of blastocoele expansion and hatching status (1–6); size and compactness of the ICM (highest score A, then B, and C); and the cohesiveness and number of TE cells (A, B, and C). We graded the ICM and TE and compared the rates of pregnancy and abortion for blastocysts of grades A, B, and C. RESULTS: The rates of pregnancy per embryo transfer cycle for ICMs of grades A, B, and C were 45.1% (829/1838), 41.7% (454/1089), and 39.4% (26/66), and abortion rates were 19.8% (164/829), 24.5% (111/454), and 26.9% (7/26) respectively. The ICM grade did not have a statistically significant effect on the rate of pregnancy or abortion. In contrast, the rates of pregnancy per embryo transfer cycle for TE grades of A, B, and C were 48.8% (488/1000), 42.4% (786/1853), and 25.0% (35/140), respectively. For grade C TE, the rate of pregnancy was significantly lower than that for TEs of grade A or B (P<0.05). The rates of abortion for TEs of grades A, B, and C were 17.8% (87/488), 23.4% (184/786), and 31.4% (11/35), respectively. Embryos with a grade C TE showed a significantly higher rate of abortion compared with those with a grade A TE (P<0.05). CONCLUSION: We found that the grade of TE was a better predictor of implantation than ICM morphology.
P-186 Tuesday, October 23, 2012 SINGLE-STEP VS. TWO-STEP MEDIA: ARE THERE ANY DIFFERENCES IN OUTCOME AND IMPLANTATION? J. Liebermann, E. J. Pelts, R. Brohammer, Y. Wagner, C. Sipe, K. Lederer. Fertility Centers of Illinois, Chicago, IL. OBJECTIVE: The culture of embryos has been approached by two culture protocols. The two-step protocol is thought to mimic the physiological interaction between specific developmental stages of the embryo within the oviduct and uterus. The opposing system in based on the ‘let the embryo choose’ principal that is analogous to a single-step culture medium. By culturing sibling oocytes in both media under the same culture conditions, we compared both groups with respect to outcome and implantation. DESIGN: Retrospective analysis. MATERIALS AND METHODS: A single-step medium (GLOBAL, IVFonline, Guilford, CT) was compared to a two-step media from SAGE (Trumbull, CT). Embryo culture was performed as group culture in 4-well dishes under oil and 5% oxygen. On day-1, following 2pn-check, sibling zygote cohorts from 181 patients (average age of 33.54.5) were randomly allocated into the SAGE or GLOBAL culture media. 953 embryos were cultured in the SAGE, whereas 1049 embryos were cultured in GLOBAL. 58 transfers from the SAGE system were compared to 123 transfers from the GLOBAL system. Clinical pregnancies showing FCA activity as determined by ultrasound. Chi square test was used for statistical analysis. RESULTS: From both culture systems an average of 1.60.5 embryos were transferred. Almost 70% of the transfers (123/181) had embryos selected from the GLOBAL culture system. Overall, the total number of embryos cryopreserved and transferred was significantly higher in the GLOBAL group (57.3%; 601/1049; P<.04) compared to the SAGE group
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ASRM Abstracts
(47.6%; 454/953). Clinical pregnancy was slightly, but not significantly higher in SAGE (58.6%; 34/58; P<.92) than in GLOBAL (56.1%; 69/ 123). Furthermore, implantation was marginal, but not significant higher in SAGE (46.1%; 42/91; P<.75) compared to GLOBAL (42%; 82/196). CONCLUSION: This trial demonstrates that embryos selected from a single-step media provide the same potential in achieving good clinical pregnancy and implantation rates compared to their sibling embryos cultured in a two-step media.
P-187 Tuesday, October 23, 2012 EFFECT OF LIF ON HUMAN EMBRYOS. A. Zhioua,a N. Mami,a S. Fourati,a H. Elloumi,a M. Khrouf,b F. Zhioua.b aCenter of Reproductive Biology, Center of Reproductive Biology, Tunis, Tunisia; bDepartment of Gynecology Obstetrics, Department of Gynecology Obstetrics, Tunis, Tunisia. OBJECTIVE: Leukemia inhibitor factor (LIF) is a cytokine with a molecular weight of 38-67KDa, which is expressed at highest concentrations in both maternal endometrial glands and extra embryonic membranes of the embryo, at about the implantation stage. The purpose of this study is to assess whether addition of LIF into a specific culture media will improve the embryo quality, susceptible to enhance pregnancy rate following embryo transfer. DESIGN: This is a blind prospective study conducted in our IVF laboratory, during the months of March and April 2012, on 338 oocytes received the day of ovum pick-ups of 50 women underwent the ICSI program. MATERIALS AND METHODS: The woman age ranges from 21 to 41 years. The oocyte cohort of each woman is randomly divided into two groups, after the microinjection: Group 1: oocytes exposed to LIF, Group 2: oocytes not exposed to LIF. Recombinant human LIF lyophilized (INVITROGEN, GIBCO) is added to microdroplets of embryo culture media (G1 Plus, VITROLIFE) at a concentration of 2.5microg/ml. These are containing the microinjected oocytes placed in culture for 2 or 3 days. We compare the embryo quality, between the exposed and the non exposed embryos to LIF at day 2 and day 3 (blastomere number, regularity and fragmentation rate). RESULTS: Exposure to LIF during the early stages of embryo development shows a statistically significant improvement in the embryo quality especially at day 3, by increasing the number of embryos with regular blastomeres (80% (group1) vs 61% (group 2), P¼0.016) and lower fragmentation rate (83% (group1) vs 59% (group 2), P¼0.003). The number of eightcell embryo is also higher in the group of embryos exposed to LIF (27% vs 17%), but with no significant difference. CONCLUSION: These results indicate that LIF could have a benefic effect on embryo quality, especially at day 3, which may increase the chance of pregnancy after ICSI. This preliminary study deserves, however, to be complemented by a larger sample in order to confirm the results obtained.
P-188 Tuesday, October 23, 2012 AN OOCYTE SPECIFIC MEDIUM FOR THE FIRST 24 HOURS OF CULTURE IMPROVES DEVELOPMENT OF GOOD QUALITY J. Stevens,b A. Janesch,b BLASTOCYSTS. R. L. Krisher,a S. D. Degelos,c G. Koulianos,c W. B. Schoolcraft.b aNational Foundation for Fertility Research, Lone Tree, CO; bColorado Center for Reproductive Medicine, Lone Tree, CO; cCenter for Reproductive Medicine, Mobile, AL. OBJECTIVE: Although oocytes have unique requirements, typically embryo culture media are used for oocyte collection and initial culture in IVF cycles. The objective of this study was to evaluate the efficacy of a medium designed specifically for the needs of oocytes, and used for the initial 24 hours of culture to support subsequent blastocyst development. DESIGN: Research study. MATERIALS AND METHODS: Oocytes (n¼575) from each patient (n¼30) were randomly allocated at retrieval to Quinns Advantage Cleavage Medium (CM, SAGE) or a new, oocyte specific medium (Oocyte Handling Medium, OHM) for the initial 24 hours of culture. ICSI was performed in OHM or CM. Fertilized embryos from both groups were then transferred for the remainder of culture to commercial sequential media (SAGE). Embryos were cultured to D5 or D6, and blastocyst development assessed. Good quality embryos were either transferred fresh (D5) or frozen (D5 or D6). Twenty four patients have undergone blastocyst transfer (19 fresh, 5 frozen).
Vol. 98, No. 3, Supplement, September 2012