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“Rescue” Intracytoplasmic Sperm Injection (ICSI) Can Result in Acceptable Pregnancy Rates in In Vitro Fertilization (IVF) Patients with Complete Fertilization Failure
were sorted into two classes, haploid versus others. A random number of cells was fixed for FISH analysis for chromosomes X, Y and 18. Results: Histopathologic analysis of the biopsy revealed generalized hypospermatogenesis. Approximately 45,000 cells were sorted from an initial group of 100,000. The incidence of haploid cells was 14.0% (n56,200), diploid was 43.1% (n519,400), and tetraploid was 26.0% (n511,700) confirming the hypospermatogenesis. Cell sorting was achieved and FISH was performed (n523), with the finding of 26.1% (6/23) Y18, 21.7% (5/23) X18 for the haploid cell group, 30.4% (7/23) diploid, and 21.7% (5/23) aneuploid cells (YY1818, XX1818, YYYXXX181818, XXX1818, and YYY181818) for the other group. Conclusions: Flow cytometric sperm analysis is a rapid and accurate technique for quantification of DNA content in azoospermic patients. Haploid cells were effectively sorted in a way that eventually may be used for ICSI. Cell sorting could also be a useful tool to optimize the time and accuracy in the diagnosis and separation of cell samples recovered from azoospermic patients.
Wednesday, October 25, 2000 4:30 P.M. O-255 A Prospective Randomized Trial of ICSI Versus IVF for the Treatment of Unexplained Infertility. A. Sertac, R. Mercan, B. Balaban, C. Alatas, S. Aksoy, B. Urman. American Hospital of Istanbul, Department of Reproductive Medicine, Istanbul, Turkey. Objectives: It has been reported that fertilization failure is common in patients with unexplained infertility undergoing conventional in-vitro fertilization (IVF). In this study we compared fertilization rates with IVF or intracytoplasmic sperm injection (ICSI) in sibling oocytes of patients with unexplained infertility. Design: Prospective randomized study. Materials and Methods: A total of 85 patients with unexplained infertility were evaluated. We included couples with normal semen parameters (ie. count, motility and morphology) and with at least 8 oocytes retrieved. Oocytes were randomly divided into two groups; conventional IVF was performed in group I while intracytoplasmic sperm injection (ICSI) was performed in group II. Results: A total of 1458 oocytes were retrieved (mean: 17.1), IVF was performed on 726 and ICSI on 722 of them. In both groups G1 and G2 media were used for incubation of the embryos. All embryo transfer procedures were performed 3 days after oocyte retrieval. As shown in table I fertilization rates were 36.6% and 83.3% in group I and II, respectively (p,0.05). Total fertilization failure was observed in sibling oocytes of 11 couples in group I, while all these couples had embryos available for transfer from their oocytes fertilized with ICSI. Cleavage rate and embryo quality were similar between the groups. Embryo quality in 5– 8 cell embryos was significantly better in ICSI group.
No of oocytes No of MII oocytes Fertilization rate Cleavage rate
IVF
ICSI
P
726
722 540 (74.8%) 83.3% 97.8%
,0.05 NS
36.5% 94.5%
Developmental stage G11G2 G31G4 G11G2 G31G4 #2 cell 5.6% 4.0% 3.7% 2.6% NS NS 2–4 cell 19.2% 13.6% 22.9% 22.9% NS ,0.05 5–8 cell 26.0% 31.6% 40.7% 25.2% ,0.05 NS
FERTILITY & STERILITYt
Conclusion: Our results show that fertilization rate is significantly higher when oocytes from patients with unexplained infertility are fertilized with ICSI. Serious consideration should be given to ICSI as the first option for in vitro fertilization in these patients.
Wednesday, October 25, 2000 4:45 P.M. O-256 “Rescue” Intracytoplasmic Sperm Injection (ICSI) Can Result in Acceptable Pregnancy Rates in In Vitro Fertilization (IVF) Patients with Complete Fertilization Failure. J. E. Nichols, T. P. Price, P. B. Miller, J. E. Johnson, H. L. Higdon, W. R. Boone. Center for Women’s Medicine, Greenville Hospital Systems, Greenville, SC. Objectives: In couples undergoing conventional IVF procedures who have had a successful oocyte retrieval, complete fertilization failure can occur in up to 10 –27% of cycles with poor fertilization rates (,35%) occurring even more frequently. Conventional reinsemination of 1 day-old unfertilized oocytes can be attempted but generally results in variable and limited fertilization and success rates. “Rescue” or 2nd-day ICSI has also been used to salvage failed or poor fertilization IVF cycles with greater than 50% fertilization rates being reported. Cleavage rates and embryo quality have been acceptable with clinical pregnancy rates of up to 7–15% and no detectable abnormalities noted in the resultant offspring. The objective of this study was to evaluate the use of rescue ICSI at our institution for salvage of complete fertilization failures after conventional IVF. Design: Retrospective chart analysis of patients who underwent IVF in our institution from 1/97 to 10/99. Data reviewed of all rescue ICSI cycles in IVF patients (non-male factor) with complete fertilization failure (n511). Materials and Methods: Non-fertilized mature oocytes from IVF cycles were micro-injected by ICSI technique 24 to 28 hours after oocyte pick-up. All embryos underwent assisted hatching prior to embryo transfer which was performed 3 or 4 days after oocyte retrieval. Fertilization and cleavage rates, polyploidy, number of embryos transferred, embryo quality, clinical pregnancy and embryo implantation rates were assessed. Results: A total of 107 metaphase II (MII) oocytes were subjected to ICSI giving rise to fertilization of 59 oocytes (55%) with a cleavage rate of 91% and polyploid (.2PN) rate of only 7%. All patients (11) went to embryo transfer with the average number of embryos replaced being 3.1 (range 1–5). Average embryo stage: 5.9 cells (range 2 cell-early morula) and grade: 2.4 (range 1– 4). Day 3 transfers were performed on 4 patients (1/4 pregnant) and Day 4 transfers in 7 (2/7 pregnant). Clinical pregnancy rate was 27% (3/11) with an implantation rate of 12%. One set of twins and two singletons, all healthy, have been delivered to date. Conclusions: Rescue ICSI can result in very acceptable pregnancy rates, comparable to the recent SART data for IVF success rates in 1996, in what would otherwise have been a failed IVF cycle without embryo transfer. Certainly, concern about the possibility of chromosomal abnormalities of 1-day old oocytes that have been reinseminated exists, however, it appears to be unfounded to date. Further studies are needed to examine other factors such as time of reinsemination, assisted hatching or day of embryo transfer that might influence the success rate of performing a rescue ICSI procedure.
S95
Wednesday, October 25, 2000 4:30 P.M. O-255 A Prospective Randomized Trial of ICSI Versus IVF for the Treatment of Unexplained Infertility. A. Sertac, R. Mercan, B. Balaban, C. Alatas, S. Aksoy, B. Urman. American Hospital of Istanbul, Department of Reproductive Medicine, Istanbul, Turkey. Objectives: It has been reported that fertilization failure is common in patients with unexplained infertility undergoing conventional in-vitro fertilization (IVF). In this study we compared fertilization rates with IVF or intracytoplasmic sperm injection (ICSI) in sibling oocytes of patients with unexplained infertility. Design: Prospective randomized study. Materials and Methods: A total of 85 patients with unexplained infertility were evaluated. We included couples with normal semen parameters (ie. count, motility and morphology) and with at least 8 oocytes retrieved. Oocytes were randomly divided into two groups; conventional IVF was performed in group I while intracytoplasmic sperm injection (ICSI) was performed in group II. Results: A total of 1458 oocytes were retrieved (mean: 17.1), IVF was performed on 726 and ICSI on 722 of them. In both groups G1 and G2 media were used for incubation of the embryos. All embryo transfer procedures were performed 3 days after oocyte retrieval. As shown in table I fertilization rates were 36.6% and 83.3% in group I and II, respectively (p,0.05). Total fertilization failure was observed in sibling oocytes of 11 couples in group I, while all these couples had embryos available for transfer from their oocytes fertilized with ICSI. Cleavage rate and embryo quality were similar between the groups. Embryo quality in 5– 8 cell embryos was significantly better in ICSI group.
No of oocytes No of MII oocytes Fertilization rate Cleavage rate
IVF
ICSI
P
726
722 540 (74.8%) 83.3% 97.8%
,0.05 NS
36.5% 94.5%
Developmental stage G11G2 G31G4 G11G2 G31G4 #2 cell 5.6% 4.0% 3.7% 2.6% NS NS 2–4 cell 19.2% 13.6% 22.9% 22.9% NS ,0.05 5–8 cell 26.0% 31.6% 40.7% 25.2% ,0.05 NS
FERTILITY & STERILITYt
Conclusion: Our results show that fertilization rate is significantly higher when oocytes from patients with unexplained infertility are fertilized with ICSI. Serious consideration should be given to ICSI as the first option for in vitro fertilization in these patients.
Wednesday, October 25, 2000 4:45 P.M. O-256 “Rescue” Intracytoplasmic Sperm Injection (ICSI) Can Result in Acceptable Pregnancy Rates in In Vitro Fertilization (IVF) Patients with Complete Fertilization Failure. J. E. Nichols, T. P. Price, P. B. Miller, J. E. Johnson, H. L. Higdon, W. R. Boone. Center for Women’s Medicine, Greenville Hospital Systems, Greenville, SC. Objectives: In couples undergoing conventional IVF procedures who have had a successful oocyte retrieval, complete fertilization failure can occur in up to 10 –27% of cycles with poor fertilization rates (,35%) occurring even more frequently. Conventional reinsemination of 1 day-old unfertilized oocytes can be attempted but generally results in variable and limited fertilization and success rates. “Rescue” or 2nd-day ICSI has also been used to salvage failed or poor fertilization IVF cycles with greater than 50% fertilization rates being reported. Cleavage rates and embryo quality have been acceptable with clinical pregnancy rates of up to 7–15% and no detectable abnormalities noted in the resultant offspring. The objective of this study was to evaluate the use of rescue ICSI at our institution for salvage of complete fertilization failures after conventional IVF. Design: Retrospective chart analysis of patients who underwent IVF in our institution from 1/97 to 10/99. Data reviewed of all rescue ICSI cycles in IVF patients (non-male factor) with complete fertilization failure (n511). Materials and Methods: Non-fertilized mature oocytes from IVF cycles were micro-injected by ICSI technique 24 to 28 hours after oocyte pick-up. All embryos underwent assisted hatching prior to embryo transfer which was performed 3 or 4 days after oocyte retrieval. Fertilization and cleavage rates, polyploidy, number of embryos transferred, embryo quality, clinical pregnancy and embryo implantation rates were assessed. Results: A total of 107 metaphase II (MII) oocytes were subjected to ICSI giving rise to fertilization of 59 oocytes (55%) with a cleavage rate of 91% and polyploid (.2PN) rate of only 7%. All patients (11) went to embryo transfer with the average number of embryos replaced being 3.1 (range 1–5). Average embryo stage: 5.9 cells (range 2 cell-early morula) and grade: 2.4 (range 1– 4). Day 3 transfers were performed on 4 patients (1/4 pregnant) and Day 4 transfers in 7 (2/7 pregnant). Clinical pregnancy rate was 27% (3/11) with an implantation rate of 12%. One set of twins and two singletons, all healthy, have been delivered to date. Conclusions: Rescue ICSI can result in very acceptable pregnancy rates, comparable to the recent SART data for IVF success rates in 1996, in what would otherwise have been a failed IVF cycle without embryo transfer. Certainly, concern about the possibility of chromosomal abnormalities of 1-day old oocytes that have been reinseminated exists, however, it appears to be unfounded to date. Further studies are needed to examine other factors such as time of reinsemination, assisted hatching or day of embryo transfer that might influence the success rate of performing a rescue ICSI procedure.
S95