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Comparison of different luteal phase support protocols for frozen embryo transfer (FET).
P-37 Comparison of different luteal phase support protocols for frozen embryo transfer (FET). J. Wilcox, J. R. Nelson, D. Potter, J. Frederick, M. Feinman, J. Batzofin. Huntington Reproductive Ctr, Pasadena, CA. Objective: To determine whether two different luteal phase hormonal support protocols result in significantly different pregnancy rates in women undergoing FET cycles. Design: Prospective randomized study. Materials/Methods: Patients were randomized into two groups for FET cycles. All patients received estradiol for follicular phase support using delestrogen 2 mg every three days for 2 doses followed by 4 mg for two weeks prior to initiation of luteal support. Standard freezing protocols were used and most embryos were frozen at 2PN. Only high-grade embryos were selected for freezing. Patients were randomized to two groups. Group-one received progesterone in oil 50 mg with Crinone 8% daily and group-two received Crinone 8% twice daily. There was no statistically significant difference in ages between the two groups. Chi-square was used for statistical analysis. Results: See table below. Luteal phase protocols. Groups
Pregnancy rates (%)
Progesterone in oil ⫹ Crinone Crinone only p value
20/43 (46.5%) 16/54 (29.6%) p ⫽ .13
Conclusions: Although this study did not reach statistical significance, there may be a selective advantage of using intramuscular progesterone in oil and Crinone. Several studies have suggested the implantation rates are lower with Crinone alone for luteal support. The combination regimen provides a physiologic replacement by providing adequate serum progesterone levels while maintaining high local concentrations in the uterine cavity. This research effort will continue to determine whether statistical significance is achieved. Supported by: Huntington Reproductive Center.
ART: LAB ISSUES P-38 Relationships between pronuclei and nucleoli morphology with the prognosis in intracytoplasmic sperm injection (ICSI) cycles. L. M. Rossi-Ferragut, A. Iaconelli Jr., C. C. Rocha, A. Medeiros, T. Aoki, E. Borges Jr. Fertility-Assisted Reproduction Ctr, Sao Paulo, Brazil. Objective: In ICSI cycles, embryos are usually selected for transfer based on the evaluation of the cleavage speed and extent of blastomere fragmentation. Several studies have shown a relationship between aspects of zygote morphology and the viability of the future embryo. Thus, these aspects have been observed with non-invasive techniques and seem to play important role in the evaluation of embryo quality. The aim of this study was to classify embryos from ICSI cycles irrespective of infertility etiology according to zygote morphology and assess the predictive value of this criterion. Rates of good embryo quality, normal cleavage and pregnancy were recorded. Design: Retrospective study from December 1999 to September 2000. Materials/Methods: Two hundred and ninety four couples (321 cycles) suffering from different types of infertility and treated by ICSI were enrolled in our study. For each embryo, the following data were recorded during sequential observations: pronuclei (PN) morphology 16 –20 hours after injection and embryo morphology about 68 hours after ICSI (day ⫹ 3). Two patterns of pronuclei morphology were observed according to the position of the PN: aligned or close pronuclei with aligned or aligning nucleoli (score 0) and separated or unequal pronuclei and scattered nucleoli (score 1). Regarding embryo morphology, embryos were classified as having good quality if more than 4 cells were present with less than 20% fragmentation at 68 hours after ICSI (day ⫹ 3). Student “t” test was employed to statistical analysis and p ⬍ 0.05 was considered statistically significant. Results: The mean maternal age (SD) of this study was 35.5 (5.5). Of 321
S124
Abstracts
ICSI cycles, we obtained 5.015 follicles, and of these 3.671 oocytes (73.2%). A number of 2.664 oocytes (72.6%) were injected and 2.343 remained intact after ICSI (63.8%). Irrespective of maternal age, in this study, 1.606 embryos (81.4% of total) were classified as an S0 (n ⫽ 743) and S1 (n ⫽ 863) in the first classification. In terms of cleavage status, the numbers found were 7% (52 S0 embryos) vs. 10,1% (87 S1 embryos) that had arrest of this process (p 0,05). Embryos selected to transfer or cryopreservation represented percentages of 86,8% S0 embryos vs. 84,5% S1 embryos (645 and 729 embryos, respectively; p 0,05). Embryo transfer occurred with both S0 and S1 embryos in 158 cycles (49.2%), with only S0 embryos in 63 cycles (19.6%) or only with S1 embryos in 69 cycles (21.5%). No embryos were transferred in 9.7% of cases (n ⫽ 31). Same results were obtained when patients were divided by maternal age (younger or older than 35 years old). Pregnancy rates per cycle and per patient were respectively, 21.7% and 23.7%, irrespective of the type of embryo transfer. Clinical pregnancy was achieved in 80% of treatment cycles in which at least one pattern 0 embryo were transferred. Conclusions: No significant differences were obtained between embryo quality and cleavage status in different patterns of embryo. Furthermore, good pronuclei and nucleoli morphology embryos seems to exhibit high probability to show better embryos on day ⫹3, based on the high percentages of pregnancy when at least one S0 embryo was transferred. Supported by: Fertility-Assisted Reproduction Center. Sa˜o Paulo, BRAZIL www.fertility.com.br/[email protected]
P-39 Embryo quality in thawed cycles from frozen pronucleated preembryos: a comparison of growth culture media. R. C. Inza, C. Bisioli, M. Tivero´n, A. Valca´rcel, M. Diradourian, C. Sueldo. Inst de Ginecologı´a y Fertilidad (IFER), Buenos Aires, Argentina. Objective: Glucose and phosphate have a detrimental effect on the first stages of embryo development, as it has been demonstrated in some species. Media lacking these two components are intended to enhance human embryo development and pregnancy rates. The purpose of the present study is to compare the reproductive performance of frozen-thawed pronucleated preembryos cultured in media with or without glucose and phosphate. Design: Retrospective and comparative study. Materials/Methods: Between September 1995 and November 2000, 263 frozen pronucleated preembryos from IVF or ICSI cycles were thawed. Human Tubal Fluid (HTF, Irvine Scientific, USA) was used as culture media containing glucose and phosphate in 157 of these cycles, while P-1 medium (Irvine Scientific, USA, without glucose and phosphate) was used in 106 cycles, both supplemented with human serum albumin (hSA) 0.5%. Preembryos remained in culture from thawing to day 3, when they were transferred. Patients from both groups, HTF and P-1, were comparable regarding infertility causes, female age, number of retrieved oocytes, proportion of viable pronucleated preembryos thawed (92.8% vs 89.9%), cleavage rate (96.2% vs 96.8%) and mean number of transferred embryos (3.6 ⫹ 1.2 vs 3.5 ⫹ 1.0). Main outcome measures were embryo quality and pregnancy and implantation rates. Statistical analysis was performed using t-test or chi square as appropriate. Results: The percentage of cycles having only good quality embryos for transfer was similar in both groups (26.9% vs 21.9%). There were significant differences regarding the overall percentage of good quality embryos in favor of the HTF group (62.9% vs 52.3%, p ⫽ 0.002). No differences were found in the pregnancy rate per ET (26.9% vs 25.7%), implantation rate per ET (10.8% vs 9.6%) or ongoing pregnancy rate per ET (22.4% vs 17.1%). Conclusions: Even though the potential detrimental effect of glucose and phosphate on the first stages of embryo development has been reported, cycles in which HTF was used as growth culture medium for thawed pronuclear preembryos showed a higher incidence of good quality embryos. This difference had no impact in achieving more pregnancies or higher implantation rate when compared with P-1 medium. The use of a free glucose and phosphate medium in this series did not give extra benefits in the analyzed parameters.
P-40 Vitamin E supplementation reduces oxidative stress and improves blastocyst development rate. X. Wang, R. K. Sharma, J. M. Goldberg, M.
Vol. 76, No. 3, Suppl. 1, September 2001
Pregnancy rates (%)
Progesterone in oil ⫹ Crinone Crinone only p value
20/43 (46.5%) 16/54 (29.6%) p ⫽ .13
Conclusions: Although this study did not reach statistical significance, there may be a selective advantage of using intramuscular progesterone in oil and Crinone. Several studies have suggested the implantation rates are lower with Crinone alone for luteal support. The combination regimen provides a physiologic replacement by providing adequate serum progesterone levels while maintaining high local concentrations in the uterine cavity. This research effort will continue to determine whether statistical significance is achieved. Supported by: Huntington Reproductive Center.
ART: LAB ISSUES P-38 Relationships between pronuclei and nucleoli morphology with the prognosis in intracytoplasmic sperm injection (ICSI) cycles. L. M. Rossi-Ferragut, A. Iaconelli Jr., C. C. Rocha, A. Medeiros, T. Aoki, E. Borges Jr. Fertility-Assisted Reproduction Ctr, Sao Paulo, Brazil. Objective: In ICSI cycles, embryos are usually selected for transfer based on the evaluation of the cleavage speed and extent of blastomere fragmentation. Several studies have shown a relationship between aspects of zygote morphology and the viability of the future embryo. Thus, these aspects have been observed with non-invasive techniques and seem to play important role in the evaluation of embryo quality. The aim of this study was to classify embryos from ICSI cycles irrespective of infertility etiology according to zygote morphology and assess the predictive value of this criterion. Rates of good embryo quality, normal cleavage and pregnancy were recorded. Design: Retrospective study from December 1999 to September 2000. Materials/Methods: Two hundred and ninety four couples (321 cycles) suffering from different types of infertility and treated by ICSI were enrolled in our study. For each embryo, the following data were recorded during sequential observations: pronuclei (PN) morphology 16 –20 hours after injection and embryo morphology about 68 hours after ICSI (day ⫹ 3). Two patterns of pronuclei morphology were observed according to the position of the PN: aligned or close pronuclei with aligned or aligning nucleoli (score 0) and separated or unequal pronuclei and scattered nucleoli (score 1). Regarding embryo morphology, embryos were classified as having good quality if more than 4 cells were present with less than 20% fragmentation at 68 hours after ICSI (day ⫹ 3). Student “t” test was employed to statistical analysis and p ⬍ 0.05 was considered statistically significant. Results: The mean maternal age (SD) of this study was 35.5 (5.5). Of 321
S124
Abstracts
ICSI cycles, we obtained 5.015 follicles, and of these 3.671 oocytes (73.2%). A number of 2.664 oocytes (72.6%) were injected and 2.343 remained intact after ICSI (63.8%). Irrespective of maternal age, in this study, 1.606 embryos (81.4% of total) were classified as an S0 (n ⫽ 743) and S1 (n ⫽ 863) in the first classification. In terms of cleavage status, the numbers found were 7% (52 S0 embryos) vs. 10,1% (87 S1 embryos) that had arrest of this process (p 0,05). Embryos selected to transfer or cryopreservation represented percentages of 86,8% S0 embryos vs. 84,5% S1 embryos (645 and 729 embryos, respectively; p 0,05). Embryo transfer occurred with both S0 and S1 embryos in 158 cycles (49.2%), with only S0 embryos in 63 cycles (19.6%) or only with S1 embryos in 69 cycles (21.5%). No embryos were transferred in 9.7% of cases (n ⫽ 31). Same results were obtained when patients were divided by maternal age (younger or older than 35 years old). Pregnancy rates per cycle and per patient were respectively, 21.7% and 23.7%, irrespective of the type of embryo transfer. Clinical pregnancy was achieved in 80% of treatment cycles in which at least one pattern 0 embryo were transferred. Conclusions: No significant differences were obtained between embryo quality and cleavage status in different patterns of embryo. Furthermore, good pronuclei and nucleoli morphology embryos seems to exhibit high probability to show better embryos on day ⫹3, based on the high percentages of pregnancy when at least one S0 embryo was transferred. Supported by: Fertility-Assisted Reproduction Center. Sa˜o Paulo, BRAZIL www.fertility.com.br/[email protected]
P-39 Embryo quality in thawed cycles from frozen pronucleated preembryos: a comparison of growth culture media. R. C. Inza, C. Bisioli, M. Tivero´n, A. Valca´rcel, M. Diradourian, C. Sueldo. Inst de Ginecologı´a y Fertilidad (IFER), Buenos Aires, Argentina. Objective: Glucose and phosphate have a detrimental effect on the first stages of embryo development, as it has been demonstrated in some species. Media lacking these two components are intended to enhance human embryo development and pregnancy rates. The purpose of the present study is to compare the reproductive performance of frozen-thawed pronucleated preembryos cultured in media with or without glucose and phosphate. Design: Retrospective and comparative study. Materials/Methods: Between September 1995 and November 2000, 263 frozen pronucleated preembryos from IVF or ICSI cycles were thawed. Human Tubal Fluid (HTF, Irvine Scientific, USA) was used as culture media containing glucose and phosphate in 157 of these cycles, while P-1 medium (Irvine Scientific, USA, without glucose and phosphate) was used in 106 cycles, both supplemented with human serum albumin (hSA) 0.5%. Preembryos remained in culture from thawing to day 3, when they were transferred. Patients from both groups, HTF and P-1, were comparable regarding infertility causes, female age, number of retrieved oocytes, proportion of viable pronucleated preembryos thawed (92.8% vs 89.9%), cleavage rate (96.2% vs 96.8%) and mean number of transferred embryos (3.6 ⫹ 1.2 vs 3.5 ⫹ 1.0). Main outcome measures were embryo quality and pregnancy and implantation rates. Statistical analysis was performed using t-test or chi square as appropriate. Results: The percentage of cycles having only good quality embryos for transfer was similar in both groups (26.9% vs 21.9%). There were significant differences regarding the overall percentage of good quality embryos in favor of the HTF group (62.9% vs 52.3%, p ⫽ 0.002). No differences were found in the pregnancy rate per ET (26.9% vs 25.7%), implantation rate per ET (10.8% vs 9.6%) or ongoing pregnancy rate per ET (22.4% vs 17.1%). Conclusions: Even though the potential detrimental effect of glucose and phosphate on the first stages of embryo development has been reported, cycles in which HTF was used as growth culture medium for thawed pronuclear preembryos showed a higher incidence of good quality embryos. This difference had no impact in achieving more pregnancies or higher implantation rate when compared with P-1 medium. The use of a free glucose and phosphate medium in this series did not give extra benefits in the analyzed parameters.
P-40 Vitamin E supplementation reduces oxidative stress and improves blastocyst development rate. X. Wang, R. K. Sharma, J. M. Goldberg, M.
Vol. 76, No. 3, Suppl. 1, September 2001