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Cumulative Conception Rates Following Intracytoplasmic Sperm Injection
Design: Prospective observational study. Materials and Methods: The results of IVF patients having a double transfer between October 1999 and January 2000 were compared with the results of IVF patients having a single day 3 embryo transfer during the same time period. Patients, for the most part having had previous implantation failure were offered a day 3 transfer if on day 2 there were multiple good quality embryos available. If on day 3 embryo development continued to be optimal and patients consented, one or two embryos were transferred. The remaining embryos were further cultured and one or two blastocysts transferred on day 5 or day 6. Results: Ten patients had a double embryo transfer and twenty-four patients underwent single embryo transfer on day 3. There was no difference in age (median534 vs 34 Median Difference (MD)51 95% Confidence Interval (CI)522–3), attempt number (median52 vs. 2 MD50 CI50 –1) or the total number of embryos transferred (median53 vs. 3 MD50 CI50 –1) between the two groups. Although the implantation (24.1% vs. 9.2% p5 0.11 CI50.8 –12.1 OR53.1) and pregnancy rates (50% vs. 32% p50.54 CI50.4 –12.3 OR52.1) were higher in the double transfer group the differences were not statistically significant. Furthermore one patient in the double transfer group who had 2 cleaved embryos and one blastocyst transferred has an ongoing triplet pregnancy with three separate placentas seen on 12 week ultrasound scan. Conclusions: The double transfer of embryos in an IVF cycle results in high pregnancy rates and the second transfer procedure does not appear to jeopardize the implantation of cleaved embryos transferred on day 3. Double transfer of embryos reduces the risk of having a complete failure to obtain blastocysts by ensuring the transfer of some embryos on day 3. These results may allow the advantages of blastocyst culture to be provided to a large group of patients since the potential disadvantage of failure to reach embryo transfer is avoided.
P-180 Cumulative Conception Rates Following Intracytoplasmic Sperm Injection. M. I. Abuzeid1, H. Fakih2, G. Abdo1, H. Salem1, M. Sasy1, I. Khan2. 1The Center for Reproductive Medicine, Hurley Medical Center, Flint, MI. 2FIRST IVF, Rochester Hills, MI. Purpose: To provide reliable prognostic information for couples seeking intracytoplasmic sperm injection (ICSI) procedure for treatment of severe male factor infertility. Design: Retrospective study. Materials and Methods: In the period between 4/94 and 12/99, 1473 consecutive ICSI cycles were performed in 1078 couples with severe male factor infertility. Mean female age was 33.8 6 5.3. Patients were divided into four groups based on female age, ,35 years, 35–37 years, 38 – 40 years, and .41 years. Tubal embryo transfer (TET) was performed in favor of uterine transfer whenever a healthy and patent fallopian tube was present. The pregnancy rate per cycle and per patient was calculated. Cumulative probabilities of achieving pregnancy (CCR) after three cycles was estimated using Kaplan-Meier method. The data was also analyzed according to the method of transfer. Results: The results are summarized in the table below:
Age ,35 35–37 38–40 .41 Total
No. of cycles
No. of patients
No. of ET@
Preg. rate/cycle
Preg. rate/patient
CCR after 3 cycles
819 284 226 144 1473
596 222 163 97 1078
4.7 6 1.7 4.6 6 1.8 4.3 6 1.9 3.7 6 2.1 4.5 6 1.8
42.2% 38% 28.8%* 18.8%** 37.1%
56.2% 45.9% 37.4%** 27.8%** 50.6%
81.3% 84.6% 68.4% 44% 78.5%
@
No. of embryos transferred by TET or uterine transfer. * P,0.005, ** P,0.001.
The pregnancy rates per cycle and per patient were 45.6% and 53.2% respectively after TET compared to 30.8% and 35.1% respectively after uterine embryo transfer, the differences were statistically significant (P,0.001). Conclusion: When the ICSI procedure is used for treatment of severe male factor infertility, women of 37 years or younger had a higher probability of achieving a pregnancy after three cycles than women of 38 years
S152
Abstracts
and older. Tubal embryo transfer appears to be more effective than uterine transfer.
P-181 Y-Bearing Enrichment of Human Spermatozoa by Hyperosmotic Centrifugation with Gel Trapping. M. T. Kan, J. J. Hariprashad, A. Akerman, Z. Rosenwaks, G. D. Palermo. The Center for Reproductive Medicine and Infertility, Weill Medical College of Cornell University, New York, NY. Objectives: Many earlier techniques for sex selection have proven to show no significant difference between X and Y-bearing spermatozoa enrichment when repeated with double-labeled sex chromosome specific DNA probes and fluorescent in-situ hybridization (FISH). Flow cytometry has proven to be the most effective technique to date, however this technique is expensive and requires the DNA to be stained with a fluorochrome in addition to laser beam excitation. The long-term effects of this procedure are not known. The purpose of this study was to attempt sex selection of Ybearing human spermatozoa with less invasive techniques. Amplification of the DNA mass difference was attempted with a hyperosmotic solution in conjunction with centrifugation and a gel to trap the spermatozoa. Design: Semen analysis samples that met normal WHO criteria were used with IRB approval. These samples were examined by triple labeled FISH to determine the percentage of X and Y-bearing spermatozoa before and after treatment. Materials and Methods: A slide for FISH was made prior treatment as a control. Four samples were washed with a routine density gradient protocol, mixed with a hyperosmotic solution, and placed in a test tube with a polyester-based gel matrix. Samples were cooled, and then centrifuged at 1,800 g for 12 minutes. The tube was gently agitated to mix the top portion of the pellet trapped in the gel. This was reconstituted and centrifuged a second time. A slide was then made for FISH. A total of 4,031 cells were counted (approximately 500 cells were counted per slide for X, Y, and chromosome 18) by one technician who was blinded to the experiment. Results: Two samples showed a significant increase in the Y-bearing spermatozoa percentage after treatment when compared to their control slides (57% vs. 49%, and 54% vs. 45%, P50.01 and P50.002 respectively). Two samples showed no significant increase in Y-bearing spermatozoa after treatment (50% vs. 46%, and 48% vs. 49%). Conclusions: Currently there is no technique which rivals flow cytometry for sex selection of spermatozoa. A less expensive and less invasive technique would be helpful for those patients who choose this option. Centrifugation in a hyperosmotic medium within a polyester-based gel tube increased the Y-bearing sperm slightly in 2 of 4 samples.
P-182 Luteal Support with Crinone 8% in 1827 Women Undergoing Assisted Reproductive Technology (ART) Procedures. H. L. Levine. Columbia Research Laboratories, Rockville Centre, New York, NY. Objectives: To assess the ability to provide luteal support using the controlled and sustained release vaginal progesterone gel, Crinone 8%, in women undergoing ART procedures. Design: Interim analysis of an open-label, multi-center trial conducted in 23 US ART centers that performed fresh IVF-ET, fresh donor IVF-ET, and controlled ovarian hyperstimulation-intrauterine insemination (IUI). Materials and Methods: Participating centers were instructed to use Crinone according to their own criteria for starting and continuing luteal support in ART cycles. Patients administered Crinone 8% (Serono Laboratories, Randolph, MA) daily. All other pertinent steps in ART treatment remained unchanged in each center. Clinical pregnancy is defined as a positive pregnancy test during weeks 6 – 8 and a gestational sac observed on ultrasound. Ongoing pregnancy is defined by identifying a fetal heartbeat at week 20. In those centers which completed at least 40 fresh IVF-ET patients, retrospective analysis also compared the clinical pregnancy rates (per retrieval) between women in the current study versus the results of women who underwent this procedure in 1997, as reported to the Society for Assisted Reproductive Technologies (SART). Weighted means were used and statistical significance was assessed using unpaired t-tests. These 1997 data from the same participating centers in this study was calculated to exclude study patients. This interim analysis included results from 1827
Vol. 74, No. 3, Suppl. 1, September 2000
P-180 Cumulative Conception Rates Following Intracytoplasmic Sperm Injection. M. I. Abuzeid1, H. Fakih2, G. Abdo1, H. Salem1, M. Sasy1, I. Khan2. 1The Center for Reproductive Medicine, Hurley Medical Center, Flint, MI. 2FIRST IVF, Rochester Hills, MI. Purpose: To provide reliable prognostic information for couples seeking intracytoplasmic sperm injection (ICSI) procedure for treatment of severe male factor infertility. Design: Retrospective study. Materials and Methods: In the period between 4/94 and 12/99, 1473 consecutive ICSI cycles were performed in 1078 couples with severe male factor infertility. Mean female age was 33.8 6 5.3. Patients were divided into four groups based on female age, ,35 years, 35–37 years, 38 – 40 years, and .41 years. Tubal embryo transfer (TET) was performed in favor of uterine transfer whenever a healthy and patent fallopian tube was present. The pregnancy rate per cycle and per patient was calculated. Cumulative probabilities of achieving pregnancy (CCR) after three cycles was estimated using Kaplan-Meier method. The data was also analyzed according to the method of transfer. Results: The results are summarized in the table below:
Age ,35 35–37 38–40 .41 Total
No. of cycles
No. of patients
No. of ET@
Preg. rate/cycle
Preg. rate/patient
CCR after 3 cycles
819 284 226 144 1473
596 222 163 97 1078
4.7 6 1.7 4.6 6 1.8 4.3 6 1.9 3.7 6 2.1 4.5 6 1.8
42.2% 38% 28.8%* 18.8%** 37.1%
56.2% 45.9% 37.4%** 27.8%** 50.6%
81.3% 84.6% 68.4% 44% 78.5%
@
No. of embryos transferred by TET or uterine transfer. * P,0.005, ** P,0.001.
The pregnancy rates per cycle and per patient were 45.6% and 53.2% respectively after TET compared to 30.8% and 35.1% respectively after uterine embryo transfer, the differences were statistically significant (P,0.001). Conclusion: When the ICSI procedure is used for treatment of severe male factor infertility, women of 37 years or younger had a higher probability of achieving a pregnancy after three cycles than women of 38 years
S152
Abstracts
and older. Tubal embryo transfer appears to be more effective than uterine transfer.
P-181 Y-Bearing Enrichment of Human Spermatozoa by Hyperosmotic Centrifugation with Gel Trapping. M. T. Kan, J. J. Hariprashad, A. Akerman, Z. Rosenwaks, G. D. Palermo. The Center for Reproductive Medicine and Infertility, Weill Medical College of Cornell University, New York, NY. Objectives: Many earlier techniques for sex selection have proven to show no significant difference between X and Y-bearing spermatozoa enrichment when repeated with double-labeled sex chromosome specific DNA probes and fluorescent in-situ hybridization (FISH). Flow cytometry has proven to be the most effective technique to date, however this technique is expensive and requires the DNA to be stained with a fluorochrome in addition to laser beam excitation. The long-term effects of this procedure are not known. The purpose of this study was to attempt sex selection of Ybearing human spermatozoa with less invasive techniques. Amplification of the DNA mass difference was attempted with a hyperosmotic solution in conjunction with centrifugation and a gel to trap the spermatozoa. Design: Semen analysis samples that met normal WHO criteria were used with IRB approval. These samples were examined by triple labeled FISH to determine the percentage of X and Y-bearing spermatozoa before and after treatment. Materials and Methods: A slide for FISH was made prior treatment as a control. Four samples were washed with a routine density gradient protocol, mixed with a hyperosmotic solution, and placed in a test tube with a polyester-based gel matrix. Samples were cooled, and then centrifuged at 1,800 g for 12 minutes. The tube was gently agitated to mix the top portion of the pellet trapped in the gel. This was reconstituted and centrifuged a second time. A slide was then made for FISH. A total of 4,031 cells were counted (approximately 500 cells were counted per slide for X, Y, and chromosome 18) by one technician who was blinded to the experiment. Results: Two samples showed a significant increase in the Y-bearing spermatozoa percentage after treatment when compared to their control slides (57% vs. 49%, and 54% vs. 45%, P50.01 and P50.002 respectively). Two samples showed no significant increase in Y-bearing spermatozoa after treatment (50% vs. 46%, and 48% vs. 49%). Conclusions: Currently there is no technique which rivals flow cytometry for sex selection of spermatozoa. A less expensive and less invasive technique would be helpful for those patients who choose this option. Centrifugation in a hyperosmotic medium within a polyester-based gel tube increased the Y-bearing sperm slightly in 2 of 4 samples.
P-182 Luteal Support with Crinone 8% in 1827 Women Undergoing Assisted Reproductive Technology (ART) Procedures. H. L. Levine. Columbia Research Laboratories, Rockville Centre, New York, NY. Objectives: To assess the ability to provide luteal support using the controlled and sustained release vaginal progesterone gel, Crinone 8%, in women undergoing ART procedures. Design: Interim analysis of an open-label, multi-center trial conducted in 23 US ART centers that performed fresh IVF-ET, fresh donor IVF-ET, and controlled ovarian hyperstimulation-intrauterine insemination (IUI). Materials and Methods: Participating centers were instructed to use Crinone according to their own criteria for starting and continuing luteal support in ART cycles. Patients administered Crinone 8% (Serono Laboratories, Randolph, MA) daily. All other pertinent steps in ART treatment remained unchanged in each center. Clinical pregnancy is defined as a positive pregnancy test during weeks 6 – 8 and a gestational sac observed on ultrasound. Ongoing pregnancy is defined by identifying a fetal heartbeat at week 20. In those centers which completed at least 40 fresh IVF-ET patients, retrospective analysis also compared the clinical pregnancy rates (per retrieval) between women in the current study versus the results of women who underwent this procedure in 1997, as reported to the Society for Assisted Reproductive Technologies (SART). Weighted means were used and statistical significance was assessed using unpaired t-tests. These 1997 data from the same participating centers in this study was calculated to exclude study patients. This interim analysis included results from 1827
Vol. 74, No. 3, Suppl. 1, September 2000