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WITHDRAWN
P-195 Tuesday, October 26, 2010 WITHDRAWN
P-196 Tuesday, October 26, 2010 MULTINUCLEATION HAS A REPETITIVE PATTERN IN PATIENTS WITH SUBSEQUENT IN VITRO FERTILIZATION (IVF) CYCLES. A. Mugica, M. Riqueros, J. M. Molina, A. Ballesteros, A. Pellicer, G. Calderon. IVI Barcelona, Barcelona, Spain; IVI Valencia, Valencia, Spain. OBJECTIVE: Multinucleation is a common feature in human embryos that has a significant negative effect on embryo implantation. This study aims to evaluate whether the pattern of embryo blastomere multinucleation (BMN) is kept in the same patient along different subsequent IVF cycles. DESIGN: Retrospective study of 152 regular patients who underwent 2 repeated cycles from 2005 to 2009 at IVI Barcelona. MATERIALS AND METHODS: A total of 1,589 embryos were graded for nuclear status on day 2 and 3 of development. Embryos with at least one multinucleated (MN) blastomere were classified as MN embryos, either on day 2 or 3. Embryo transfers were performed on day 3. According to the percentage of MN embryos, patients were classified in the following categories. GroupA: 0-20% MN embryos; GroupB: 21-49%; GroupC: 50100%. c2-test for trend was used for statistic analysis. Values are reported as means SD or as percentages. RESULTS: There are not significant differences between the two subsequent cycles regarding maternal age (1: 34.59 3.38 vs 2: 35.34 3.34) and the percentage of MN embryos (Day 2: 221 (28.37%) vs 201 (24.81%); Day 3: 141 (18.10%) vs 159 (19.63%)). We observed a significant positive relationship between the two cycles in terms of the percentage of MN embryos per cycle (Table 1).
TABLE1. Distribution of patients A No. of patients 1st Cycle (%) 76 (50.00) with MN on day 2 2nd Cycle (%) 84 (55.26) No. of patients 1st Cycle (%) 108 (71.05) with MN on day 3 2nd Cycle (%) 93 (61.18)
B
C
P-Value
37 (24.34) 39 (25.66) 32 (21.05) 36 (23.69) 23 (15.13) 21 (13.82)
0.003
28 (18.42) 31 (20.40)
0.04
CONCLUSION: The origin of embryo BMN still remains unknown and it most probably appears as a result of combined intrinsic and extrinsic factors. According to our data, the presence of MN embryos appears to be an intrinsic characteristic of the patient, which repeats in subsequent IVF cycles. Thus, oocyte quality has an important role in the appearance of embryo BMN.
MATERIALS AND METHODS: Images in 7 focal planes of each embryo were acquired every 20 min during a 70 hr incubation for cycles in the ES. All 356 treatment cycles (SI: N¼279; ES: N¼77) were conducted at IVI during the period 10/8/2009 – 15/3/2010. The inclusion criteria were cycles with: transfer of embryos, ICSI, and >3 aspirated oocytes. Treatments with PGD were excluded. Ongoing pregnancy (OP) was evaluated as biochemical pregnancy (BQ) and subsequent observation of gestational sac(s). Fisher’s exact two-sided test was used on data in contingency tables. RESULTS: The BQ rates were SI: 59%, ES: 64%, p¼0.51, and the OP rates evaluated were SI: 49%, ES: 55%, p¼0.44. Both rates were slightly higher for ES than for SI cycles but the difference was not significant. Factors related to outcome were tested with no significant difference between the SI and ES cycles i.e. age, frequency of abortions, # transferred + frozen embryos, aspirated oocytes, transfer day, proportion of single/double/triple transfers, doses of FSH, LH and hMG and proportion of agonist/antagonist/other treatments. CONCLUSION: No significant difference was found between the rates of OP in the SI and ES. Although the number of treatments in the ES is limited (N¼77), the data corroborate that the ES provides as good opportunities for embryo development as the SI, and without adverse effect of image acquisition. Future studies must evaluate, if the additional information about cell division patterns can improve embryo selection.
P-198 Tuesday, October 26, 2010 EMBRYO DEVELOPMENT AND PREGNANCY RATES OF IN VITRO PRODUCED BOVINE EMBRYOS CULTURED IN HIGH AND LOW-OXYGEN (O2) ATMOSPHERE TENSION WITH SEQUENTIAL MEDIA (SM). P. Koyner, J. Pino, R. Berbey. Animal Reproduction and Genetic Research Center, David, Chiriqui, Panama; Centro Fecundar, Panama, Panama. OBJECTIVE: To evaluate embryo development and pregnancy rates with two O2 concentrations in bovine embryo culture, using SM. DESIGN: Randomized trial. MATERIALS AND METHODS: Oocytes collected from slaughterhouse ovaries cows were matured and fertilized in vitro using standard procedures. The resultants zygotes were cultured in a media 1 plus bovine serum albumin-fatty acid free (3 mg/ml) for 96 hours post-insemination (hpi) followed by a media 2 for 48 hours additional culture (Koyner et al., 2010), at 38.5 C in either an atmosphere of 6% CO2 and 20% O2 (treatment 1; n¼ 500) or an atmosphere of 6% CO2, 5%O2 and 89% N2 (treatment 2; n¼ 550). A subset of quality grade 1 blastocysts produced in treatment 1 (n¼ 25) or treatment 2 (n¼ 25) were transferred nonsurgically to synchronized recipients. The experiment was replicated four times and data were analyzed using chi-square. RESULTS: There was no significant differences during embryo development at 48, 96 and 144 hpi (see table 1) between treatment groups. Following transfers, pregnancy rates at 60 days were similar between blastocysts produced in treatment 1 and those produced in treatment 2 (33% vs. 37%; respectively).
EMBRYO CULTURE P-197 Tuesday, October 26, 2010 PREGNANCY RATES AFTER INCUBATION IN NEW TIME-LAPSE INCUBATOR (EMBRYOSCOPE) PROVIDING DETAILED INFORMATION ABOUT EMBRYO DEVELOPMENT COMPARED TO INCUBATION IN A STANDARD INCUBATOR. M. Meseguer, K. M. Hilligsøe, K. S. Pedersen, J. Herrero, A. Tejera, N. Garrido. Clinical Embryology Laboratory, IVI Valencia, Valencia, Spain; Unisense Fertilitech, Aarhus, Aarhus N, Denmark. OBJECTIVE: A new time-lapse incubator for embryo development, the EmbryoScope (ES) gives determines e.g. cell division patterns. Our goal was to evaluate if the ES provides an environment for embryo development of similar quality as the standard incubator (SI) by comparing the pregnancy rates for treatments with the SI or the ES. Embryo selection was in both cases based on observations at a few discrete time-points. Additional information provided by the ES was not used to improve selection. DESIGN: Paired cohorts retrospective study.
S150
Abstracts
TABLE 1. Effect of O2 tension during bovine embryo culture with SM.
Treatment 1 2
Zygotes (n)
R2 cells; 48hpi (%)
R8 cells; 96 hpi (%)
Blastocysts; 144 hpi (%)
500 550
420 (84%) 468 (85%)
272 (54%) 330 (60%)
191 (38%) 242 (44%)
No significant differences. CONCLUSION: These results demonstrate that SM employed in this study, which contain antioxidants and growth factors, can support in vitro embryo development and his viability under atmosphere of high O2 tension in a similar way to atmosphere of low O2 tension. Additional transfers are needed to determine if the use of this SM will avoid applying low O2 tension during bovine culture, which could be important regarding high costs of this system. Supported by: Oteima University. Special thanks are extended to Batipa Farm and Dr. Reynaldo de Armas.
Vol. 94., No. 4, Supplement, September 2010
P-196 Tuesday, October 26, 2010 MULTINUCLEATION HAS A REPETITIVE PATTERN IN PATIENTS WITH SUBSEQUENT IN VITRO FERTILIZATION (IVF) CYCLES. A. Mugica, M. Riqueros, J. M. Molina, A. Ballesteros, A. Pellicer, G. Calderon. IVI Barcelona, Barcelona, Spain; IVI Valencia, Valencia, Spain. OBJECTIVE: Multinucleation is a common feature in human embryos that has a significant negative effect on embryo implantation. This study aims to evaluate whether the pattern of embryo blastomere multinucleation (BMN) is kept in the same patient along different subsequent IVF cycles. DESIGN: Retrospective study of 152 regular patients who underwent 2 repeated cycles from 2005 to 2009 at IVI Barcelona. MATERIALS AND METHODS: A total of 1,589 embryos were graded for nuclear status on day 2 and 3 of development. Embryos with at least one multinucleated (MN) blastomere were classified as MN embryos, either on day 2 or 3. Embryo transfers were performed on day 3. According to the percentage of MN embryos, patients were classified in the following categories. GroupA: 0-20% MN embryos; GroupB: 21-49%; GroupC: 50100%. c2-test for trend was used for statistic analysis. Values are reported as means SD or as percentages. RESULTS: There are not significant differences between the two subsequent cycles regarding maternal age (1: 34.59 3.38 vs 2: 35.34 3.34) and the percentage of MN embryos (Day 2: 221 (28.37%) vs 201 (24.81%); Day 3: 141 (18.10%) vs 159 (19.63%)). We observed a significant positive relationship between the two cycles in terms of the percentage of MN embryos per cycle (Table 1).
TABLE1. Distribution of patients A No. of patients 1st Cycle (%) 76 (50.00) with MN on day 2 2nd Cycle (%) 84 (55.26) No. of patients 1st Cycle (%) 108 (71.05) with MN on day 3 2nd Cycle (%) 93 (61.18)
B
C
P-Value
37 (24.34) 39 (25.66) 32 (21.05) 36 (23.69) 23 (15.13) 21 (13.82)
0.003
28 (18.42) 31 (20.40)
0.04
CONCLUSION: The origin of embryo BMN still remains unknown and it most probably appears as a result of combined intrinsic and extrinsic factors. According to our data, the presence of MN embryos appears to be an intrinsic characteristic of the patient, which repeats in subsequent IVF cycles. Thus, oocyte quality has an important role in the appearance of embryo BMN.
MATERIALS AND METHODS: Images in 7 focal planes of each embryo were acquired every 20 min during a 70 hr incubation for cycles in the ES. All 356 treatment cycles (SI: N¼279; ES: N¼77) were conducted at IVI during the period 10/8/2009 – 15/3/2010. The inclusion criteria were cycles with: transfer of embryos, ICSI, and >3 aspirated oocytes. Treatments with PGD were excluded. Ongoing pregnancy (OP) was evaluated as biochemical pregnancy (BQ) and subsequent observation of gestational sac(s). Fisher’s exact two-sided test was used on data in contingency tables. RESULTS: The BQ rates were SI: 59%, ES: 64%, p¼0.51, and the OP rates evaluated were SI: 49%, ES: 55%, p¼0.44. Both rates were slightly higher for ES than for SI cycles but the difference was not significant. Factors related to outcome were tested with no significant difference between the SI and ES cycles i.e. age, frequency of abortions, # transferred + frozen embryos, aspirated oocytes, transfer day, proportion of single/double/triple transfers, doses of FSH, LH and hMG and proportion of agonist/antagonist/other treatments. CONCLUSION: No significant difference was found between the rates of OP in the SI and ES. Although the number of treatments in the ES is limited (N¼77), the data corroborate that the ES provides as good opportunities for embryo development as the SI, and without adverse effect of image acquisition. Future studies must evaluate, if the additional information about cell division patterns can improve embryo selection.
P-198 Tuesday, October 26, 2010 EMBRYO DEVELOPMENT AND PREGNANCY RATES OF IN VITRO PRODUCED BOVINE EMBRYOS CULTURED IN HIGH AND LOW-OXYGEN (O2) ATMOSPHERE TENSION WITH SEQUENTIAL MEDIA (SM). P. Koyner, J. Pino, R. Berbey. Animal Reproduction and Genetic Research Center, David, Chiriqui, Panama; Centro Fecundar, Panama, Panama. OBJECTIVE: To evaluate embryo development and pregnancy rates with two O2 concentrations in bovine embryo culture, using SM. DESIGN: Randomized trial. MATERIALS AND METHODS: Oocytes collected from slaughterhouse ovaries cows were matured and fertilized in vitro using standard procedures. The resultants zygotes were cultured in a media 1 plus bovine serum albumin-fatty acid free (3 mg/ml) for 96 hours post-insemination (hpi) followed by a media 2 for 48 hours additional culture (Koyner et al., 2010), at 38.5 C in either an atmosphere of 6% CO2 and 20% O2 (treatment 1; n¼ 500) or an atmosphere of 6% CO2, 5%O2 and 89% N2 (treatment 2; n¼ 550). A subset of quality grade 1 blastocysts produced in treatment 1 (n¼ 25) or treatment 2 (n¼ 25) were transferred nonsurgically to synchronized recipients. The experiment was replicated four times and data were analyzed using chi-square. RESULTS: There was no significant differences during embryo development at 48, 96 and 144 hpi (see table 1) between treatment groups. Following transfers, pregnancy rates at 60 days were similar between blastocysts produced in treatment 1 and those produced in treatment 2 (33% vs. 37%; respectively).
EMBRYO CULTURE P-197 Tuesday, October 26, 2010 PREGNANCY RATES AFTER INCUBATION IN NEW TIME-LAPSE INCUBATOR (EMBRYOSCOPE) PROVIDING DETAILED INFORMATION ABOUT EMBRYO DEVELOPMENT COMPARED TO INCUBATION IN A STANDARD INCUBATOR. M. Meseguer, K. M. Hilligsøe, K. S. Pedersen, J. Herrero, A. Tejera, N. Garrido. Clinical Embryology Laboratory, IVI Valencia, Valencia, Spain; Unisense Fertilitech, Aarhus, Aarhus N, Denmark. OBJECTIVE: A new time-lapse incubator for embryo development, the EmbryoScope (ES) gives determines e.g. cell division patterns. Our goal was to evaluate if the ES provides an environment for embryo development of similar quality as the standard incubator (SI) by comparing the pregnancy rates for treatments with the SI or the ES. Embryo selection was in both cases based on observations at a few discrete time-points. Additional information provided by the ES was not used to improve selection. DESIGN: Paired cohorts retrospective study.
S150
Abstracts
TABLE 1. Effect of O2 tension during bovine embryo culture with SM.
Treatment 1 2
Zygotes (n)
R2 cells; 48hpi (%)
R8 cells; 96 hpi (%)
Blastocysts; 144 hpi (%)
500 550
420 (84%) 468 (85%)
272 (54%) 330 (60%)
191 (38%) 242 (44%)
No significant differences. CONCLUSION: These results demonstrate that SM employed in this study, which contain antioxidants and growth factors, can support in vitro embryo development and his viability under atmosphere of high O2 tension in a similar way to atmosphere of low O2 tension. Additional transfers are needed to determine if the use of this SM will avoid applying low O2 tension during bovine culture, which could be important regarding high costs of this system. Supported by: Oteima University. Special thanks are extended to Batipa Farm and Dr. Reynaldo de Armas.
Vol. 94., No. 4, Supplement, September 2010