- Email: [email protected]
Is aneuploidy screening with fluorescent in situ hybridization (FISH) a better selection criteria for human embryo transfer than embryo morphology?
produce euploid embryos may be a result of defects resulting from in vitro maturation of the oocyte. It is also possible the aged sperm will not exhibit the ability to form a normal sperm aster at fertilization. Supported by: None
Wednesday, October 20, 2004 3:00 P.M. O-243 Sperm cell aneuploidy in semen, swim-up preparations and the zona pellucida of ova and embryos from in vitro fertilization. S. B. Hudson, S. M. Dobin, K. J. Clawson, T. J. Wincek, J. F. Pliego, T. J. Kuehl. Scott & White, Temple, TX. OBJECTIVE: Previously we demonstrated that sperm cells archived within the zona pellucida (ZP) and those within a swim-up preparation demonstrate improved morphology compared to those in semen. This study evaluates the incidence of aneuploidy of chromosomes 13, 18, 21, X and Y and compares the incidence in semen, the inseminated populations, and those archived within the ZP using fluorescent in situ hybridization (FISH). This study tests the hypothesis that a subset of sperm cells with lower incidence of aneuploidy is selected. DESIGN: FISH analysis of sperm cells in semen, swim-up preparations, and the ZP of discarded ova or degenerating embryos produced during in vitro fertilization (IVF). MATERIALS AND METHODS: Anonymous samples of sperm cells from 10 original ejaculates and paired swim-up preparations, and ZP from couples undergoing IVF were obtained. Sperm cells archived in ZP were harvested by mechanical washing and enzymatic digestion. Sperm cells processed to expand the nucleus were labeled using a commercial FISH protocol. Slides were evaluated with a fluorescence microscope equipped for dual-color detection with a triple bandpass filter. An average 831 sperm cells were evaluated from each preparation for chromosomes 18, X and Y and an average 565 sperm cells were evaluated for chromosomes 21 and 13. Results were compared using analysis of variance for repeated measures. RESULTS: As in previous report, sperm cell motility and morphology were significantly (p⬍0.0001) improved in the swim-up preparations. The proportions of sperm cells with a normal haploid complement of chromosomes 18 and X or Y were not different (p⫽0.22) between semen (95.5 ⫾ 0.6%; mean ⫾ SE) and paired swim-up (96.5 ⫾ 0.8%) preparations. In preliminary results of ZP sperm, a normal complement of 18 and X or Y was observed in 94% (80 to 99%; 95% CI). No incidence of disomy was noted within the ZP (N ⫽ 35). Disomy rates of chromosomes 18, 13, 21 and X or Y within semen populations were 0.6 ⫾ 0.1%, 0.5 ⫾ 0.2%, 0.5 ⫾ 0.1% and 1.2 ⫾ 0.2%, respectively; and within swim-up populations were 0.2 ⫾ 0.1%, 0.4 ⫾ 0.02%, 0.5 ⫾ 0.3% and 0.7 ⫾ 0.1%, respectively. Nullisomy rates of chromosomes 13, 18, 21 and X or Y within semen populations were 0.7 ⫾ 0.2%, 1.0 ⫾ 0.2%, 0.7 ⫾ 0.3% and 1.6 ⫾ 0.3%, respectively; and within swim-up populations were 1.3 ⫾ 0.7%, 1.9 ⫾ 1.3%, 3.2 ⫾ 2.6% and 1.4 ⫾ 0.2%, respectively. Our findings of no difference in aneuploidy of chromosomes 13, 18, 21, X and Y for swim-up versus semen populations are in concordance with those demonstrated by VanDyk et al. in 2000 but disagree with the findings of others (Li et al., 1998; Ong et al., 2002; Jakab et al., 2003) within recent years. CONCLUSION: Our data through previous studies suggests that selection for morphology and motility takes place in the swim-up preparation and the ZP. This study indicates selection does not appear to alter the frequency of sperm cell chromosomal abnormalities compatible with life. Supported by: Noble Centennial Endowment
Wednesday, October 20, 2004 4:45 P.M. O-244 A comparative study on nuclear DNA integrity and morphology of human spermatozoa processed by three different methods. A. C. Varghese, A. K. Bhattacharyya, H. Roy, S. S. Allamaneni, J. Bhattacharya, A. Agarwal. University of Calcutta, Calcutta, India; Shivam Hospital & NHC, Kankarbagh, India; Cleveland Clinic Foundation, Cleveland, OH; A.H. IVF & Infertility Research Center, Calcutta, India.
S98
Abstracts
OBJECTIVE: Various sperm preparation methods are in use to isolate best quality sperm for use in assisted reproductive technology programs. The swim up procedure either from neat semen or from a washed pellet and density gradient centrifugation (DGC) is the most commonly used techniques to enrich spermatozoa. Our objective was to study the effects of DGC and swim up techniques on nuclear DNA integrity and morphology of human spermatozoa. We also evaluated the efficacy of swim up method after DGC in isolating spermatozoa with chromatin integrity and normal morphology. DESIGN: Prospective study. MATERIALS AND METHODS: Twenty men attending an assisted reproduction laboratory for female factor infertility were included in this study. Ejaculates were divided into three aliquots. Sperm DNA normality was assessed by acridine orange (AO) fluorescence and morphology by Pap staining method, both before and after 1) double-layered DGC (PureSperm40% and 80%), 2) direct swim up from neat semen, 3) swim up from washed pellet and 4) swim up using pellet from DGC. The AO status of spermatozoa in 40% fraction was also assessed. RESULTS: Following DGC and swim up procedures, the percentage of nuclear maturity increased significantly compared to the neat semen (89.91⫾6.58) (Table). The nuclear maturity values (mean⫾SD) were highest after swim up from 80% fraction of DGC (97.08⫾3.93). A nuclear maturity value of 87.35⫾6.89 in 40% fraction of density gradient was significantly lower compared to neat semen (P ⫽ 0.0118). In addition, there was a significant improvement in morphologically normal forms after the different processing techniques (Table). Best enrichment of normal morphology spermatozoa noted after swim-up procedure from 80% fraction of DGC. CONCLUSION: The results indicate that a swim up step, following density gradient centrifugation, can significantly improve the percentage of spermatozoa with normal nuclear maturity and morphology compared to neat semen sample and other sperm processing techniques. Although, a high yield of morphologically normal forms can be separated after a swim up from washed pellet, the integrity of sperm DNA may be compromised due to the high levels of reactive oxygen species generated during washing procedure. This is especially relevant if the sample contain high levels of leukocytes and immature spermatozoa. Supported by: None
Wednesday, October 20, 2004 5:00 P.M. O-245 Is aneuploidy screening with fluorescent in situ hybridization (FISH) a better selection criteria for human embryo transfer than embryo morphology? H. Kim, J. F. Mayer, C. Reilly, P. W. Zarutskie. Mission Reproductive Center, Laguna Niguel, CA. OBJECTIVE: IVF centers select embryos for transfer according to morphology. This study was undertaken to understand the relationship between chromosome abnormalities and embryo morphology and to evaluate the impact of preimplantation genetic diagnosis (PGD) screening on ART outcomes. DESIGN: Retrospective analysis of PGD cycles investigated for aneuploidy from 11/02 to 3/04. MATERIALS AND METHODS: A total of 420 embryos were analyzed for 49 patients. These were from 45 fresh cycles and 10 frozen cycles. Mean age was 37.5 ⫾5.4 and included 24 patients (49%) who were ⱖ40 years old. Embryo morphology were recorded on day 3 and 5. Day 3 embryos, were
Vol. 82, Suppl. 2, September 2004
divided into 3 groups (Grades A: good; B: intermediate; C: poor) based upon standard criteria: number of cells, blastomere size and % cytoplasmic fragmentation. Day 5 embryos were divided into 4 groups (Good blastocyst; Poor blastocyst; Morula; Arrested embryo) based on cell number and morphological quality of the inner cell mass and trophectoderm. Blastomere biopsy was performed on day 3 using standard procedures. FISH using nine probes for chromosomes X, Y, 13, 15, 16, 17, 18, 21 and 22 was applied for aneuploidy screening. Embryo transfer was carried out only if the cycle had normal embryo(s). RESULTS: Chromosomal defects were found in 74% of all Day 3 embryos. There was a trend towards increasing rate of abnormality with poorer morphology (A: 63%; B: 78%; C: 92%). A similar pattern of increased chromosomal abnormalities was also observed with the slower embryo cleavage rates (Table 1). Aneuploidy was found in 73% of the day 5 embryos. Table 2 shows chromosomal abnormality in 42 % of Good blastocysts and 54 % of Poor blastocysts. Twelve cycles (21.8%) out of a total 55 cycles could not be transferred because all of the tested embryos were identified as aneuploid embryos. Mean number of transferred embryos was 1.7 and clinical pregnancy rates in aged (ⱖ 40 years) and younger (⬍ 40 years) patients were 50 % (10/20) and 52 % (12/23) per transfer (or 42% and 48% per cycle), respectively. CONCLUSION: This study suggests embryo morphology correlates only crudely with chromosomal abnormality. Using morphology or cell number failed to separate out a normal embryo population. We conclude that PGD aneuploidy screening may be an alternate way of selecting embryos for transfer. Using aneuploidy screening, we have been able to achieve elevated pregnancy rates in the older patient population. These results support further investigating the use of aneploidy screening for embryo selection. Supported by: None
were then transferred to one of the following culture media: a) Vitro Life Series: IVF500, n⫽4,248; G1.2, n⫽7,323; G1.3, n⫽5,329; or b) Irvine Scientific: P1, n⫽10,805. Embryos were stratified according to growth medium used and patient age (⬍35y and ⱖ35y), and evaluated for development to the ⱖ 8-cell stage. Data were analyzed using Chi square with p⬍0.05 considered statistically significant. RESULTS: Media type significantly impacted development of embryos to ⱖ 8-cell stage (IVF500: 27.7%; G1.2: 34.7%; G1.3: 38.7%; P1: 43.3%; p⬍0.0001). Of note, there was a significant improvement in the % of embryos at ⱖ8-cells as each new generation of Vitro Life medium was introduced (p⬍0.0001). As shown in the table, within age group, medium type continued to impact development to at least the 8-cell stage (p⬍0.0001 for both age groups). However, this impact was not consistently observed within across age groups and within medium.
CONCLUSION: Our data indicate that cleavage rate is influenced by patient age and culture medium type. This relationship should be taken into consideration when establishing quality assurance target values. For example, our current day 5 inclusion criteria were developed using the IVF500 system. Given our current medium system (G1.3) and the results of this analysis, we must now revise our Day 5 inclusion criteria because the relevant target values have changed. Constant surveillance of relevant target values must be maintained in the IVF laboratory since improvements are continually being made in ART systems as demonstrated by the improvement across the generations of Vitro Life media. Supported by: None.
Wednesday, October 20, 2004 5:30 P.M. Wednesday, October 20, 2004 5:15 P.M. O-246 Variables that impact the use of cleavage rates in quality assurance assessment. S. L. Benedict, K. V. Jackson, K. Kelleher, Y. Pan, A. Nureddin, C. Racowsky. Brigham and Women’s Hospital, Harvard Medical School, Boston, MA. OBJECTIVE: Numerous studies have shown that embryo morphology plays a key role in IVF outcome. Development to at least the 8-cell stage has been correlated with improved implantation potential, thereby providing a potentially useful marker for evaluating laboratory performance. However, it is essential to identify what variables impact embryo cleavage when using this type of marker for quality assessment so that relevant target values are identified for each technique being assessed. This study was undertaken to test the hypothesis that culture media type and patient age both influence development of the embryo to at least the 8-cell stage and therefore must be taken into consideration when assessing laboratory performance using ⱖ 8-cell embryos as a marker. DESIGN: Retrospective data analysis. MATERIALS AND METHODS: 7,030 ART cycles performed in our institution from January 1, 1998 to December 31, 2003 were retrospectively reviewed. The study dataset included 3,678 IVF cycles with or without assisted hatching. Oocytes were inseminated in Ham’s F10⫹5% HSA or 10% SSS and evaluated 16 –18 hr later for PN formation. Diploid zygotes
FERTILITY & STERILITY威
O-247 Oolema breakage after ICSI: A technical problem or oocyte quality? I. Aboujaoude, H. Maroun, M. Hamze, F. Hachem, I. Alam, P. Saroufim. Center for Reproductive Medicine and Genetics.Aboujaoude Hospital, Jaleldib, Lebanon. OBJECTIVE: During ICSI the oolema can break and results in oocyte lysis. Oolema breakage after ICSI is underreported and is attributed to a technical difficulty during ICSI procedure or an abnormal pipette diameter. Very few reports have analyzed oolema breakage from oocyte quality. DESIGN: We compared ICSI results (age, length of stimulation, estradiol level, pregnancy rate, miscarriage rate) according to oolema breakage. MATERIALS AND METHODS: 471 couples underwent ICSI cycles after controlled ovarian hyperstimulation. The ICSI procedures were performed by a single technician using Cook® pipette. Group A have less than 15% of oocytes breakage after ICSI when evaluated on day1 and group B have 15% or more oocyte breakage after ICSI. RESULTS: The results are summarized in table 1. In group A smaller number of ampoules was used for ovulation induction than group B. In group A we have less oocytes retrieved and smaller number of immature oocytes. In group A more embryo were obtained than group B. No statistical differences were found between group A and B in terms of implantation rates, pregnancy rates and miscarriage rates. Results table 1
S99
Wednesday, October 20, 2004 3:00 P.M. O-243 Sperm cell aneuploidy in semen, swim-up preparations and the zona pellucida of ova and embryos from in vitro fertilization. S. B. Hudson, S. M. Dobin, K. J. Clawson, T. J. Wincek, J. F. Pliego, T. J. Kuehl. Scott & White, Temple, TX. OBJECTIVE: Previously we demonstrated that sperm cells archived within the zona pellucida (ZP) and those within a swim-up preparation demonstrate improved morphology compared to those in semen. This study evaluates the incidence of aneuploidy of chromosomes 13, 18, 21, X and Y and compares the incidence in semen, the inseminated populations, and those archived within the ZP using fluorescent in situ hybridization (FISH). This study tests the hypothesis that a subset of sperm cells with lower incidence of aneuploidy is selected. DESIGN: FISH analysis of sperm cells in semen, swim-up preparations, and the ZP of discarded ova or degenerating embryos produced during in vitro fertilization (IVF). MATERIALS AND METHODS: Anonymous samples of sperm cells from 10 original ejaculates and paired swim-up preparations, and ZP from couples undergoing IVF were obtained. Sperm cells archived in ZP were harvested by mechanical washing and enzymatic digestion. Sperm cells processed to expand the nucleus were labeled using a commercial FISH protocol. Slides were evaluated with a fluorescence microscope equipped for dual-color detection with a triple bandpass filter. An average 831 sperm cells were evaluated from each preparation for chromosomes 18, X and Y and an average 565 sperm cells were evaluated for chromosomes 21 and 13. Results were compared using analysis of variance for repeated measures. RESULTS: As in previous report, sperm cell motility and morphology were significantly (p⬍0.0001) improved in the swim-up preparations. The proportions of sperm cells with a normal haploid complement of chromosomes 18 and X or Y were not different (p⫽0.22) between semen (95.5 ⫾ 0.6%; mean ⫾ SE) and paired swim-up (96.5 ⫾ 0.8%) preparations. In preliminary results of ZP sperm, a normal complement of 18 and X or Y was observed in 94% (80 to 99%; 95% CI). No incidence of disomy was noted within the ZP (N ⫽ 35). Disomy rates of chromosomes 18, 13, 21 and X or Y within semen populations were 0.6 ⫾ 0.1%, 0.5 ⫾ 0.2%, 0.5 ⫾ 0.1% and 1.2 ⫾ 0.2%, respectively; and within swim-up populations were 0.2 ⫾ 0.1%, 0.4 ⫾ 0.02%, 0.5 ⫾ 0.3% and 0.7 ⫾ 0.1%, respectively. Nullisomy rates of chromosomes 13, 18, 21 and X or Y within semen populations were 0.7 ⫾ 0.2%, 1.0 ⫾ 0.2%, 0.7 ⫾ 0.3% and 1.6 ⫾ 0.3%, respectively; and within swim-up populations were 1.3 ⫾ 0.7%, 1.9 ⫾ 1.3%, 3.2 ⫾ 2.6% and 1.4 ⫾ 0.2%, respectively. Our findings of no difference in aneuploidy of chromosomes 13, 18, 21, X and Y for swim-up versus semen populations are in concordance with those demonstrated by VanDyk et al. in 2000 but disagree with the findings of others (Li et al., 1998; Ong et al., 2002; Jakab et al., 2003) within recent years. CONCLUSION: Our data through previous studies suggests that selection for morphology and motility takes place in the swim-up preparation and the ZP. This study indicates selection does not appear to alter the frequency of sperm cell chromosomal abnormalities compatible with life. Supported by: Noble Centennial Endowment
Wednesday, October 20, 2004 4:45 P.M. O-244 A comparative study on nuclear DNA integrity and morphology of human spermatozoa processed by three different methods. A. C. Varghese, A. K. Bhattacharyya, H. Roy, S. S. Allamaneni, J. Bhattacharya, A. Agarwal. University of Calcutta, Calcutta, India; Shivam Hospital & NHC, Kankarbagh, India; Cleveland Clinic Foundation, Cleveland, OH; A.H. IVF & Infertility Research Center, Calcutta, India.
S98
Abstracts
OBJECTIVE: Various sperm preparation methods are in use to isolate best quality sperm for use in assisted reproductive technology programs. The swim up procedure either from neat semen or from a washed pellet and density gradient centrifugation (DGC) is the most commonly used techniques to enrich spermatozoa. Our objective was to study the effects of DGC and swim up techniques on nuclear DNA integrity and morphology of human spermatozoa. We also evaluated the efficacy of swim up method after DGC in isolating spermatozoa with chromatin integrity and normal morphology. DESIGN: Prospective study. MATERIALS AND METHODS: Twenty men attending an assisted reproduction laboratory for female factor infertility were included in this study. Ejaculates were divided into three aliquots. Sperm DNA normality was assessed by acridine orange (AO) fluorescence and morphology by Pap staining method, both before and after 1) double-layered DGC (PureSperm40% and 80%), 2) direct swim up from neat semen, 3) swim up from washed pellet and 4) swim up using pellet from DGC. The AO status of spermatozoa in 40% fraction was also assessed. RESULTS: Following DGC and swim up procedures, the percentage of nuclear maturity increased significantly compared to the neat semen (89.91⫾6.58) (Table). The nuclear maturity values (mean⫾SD) were highest after swim up from 80% fraction of DGC (97.08⫾3.93). A nuclear maturity value of 87.35⫾6.89 in 40% fraction of density gradient was significantly lower compared to neat semen (P ⫽ 0.0118). In addition, there was a significant improvement in morphologically normal forms after the different processing techniques (Table). Best enrichment of normal morphology spermatozoa noted after swim-up procedure from 80% fraction of DGC. CONCLUSION: The results indicate that a swim up step, following density gradient centrifugation, can significantly improve the percentage of spermatozoa with normal nuclear maturity and morphology compared to neat semen sample and other sperm processing techniques. Although, a high yield of morphologically normal forms can be separated after a swim up from washed pellet, the integrity of sperm DNA may be compromised due to the high levels of reactive oxygen species generated during washing procedure. This is especially relevant if the sample contain high levels of leukocytes and immature spermatozoa. Supported by: None
Wednesday, October 20, 2004 5:00 P.M. O-245 Is aneuploidy screening with fluorescent in situ hybridization (FISH) a better selection criteria for human embryo transfer than embryo morphology? H. Kim, J. F. Mayer, C. Reilly, P. W. Zarutskie. Mission Reproductive Center, Laguna Niguel, CA. OBJECTIVE: IVF centers select embryos for transfer according to morphology. This study was undertaken to understand the relationship between chromosome abnormalities and embryo morphology and to evaluate the impact of preimplantation genetic diagnosis (PGD) screening on ART outcomes. DESIGN: Retrospective analysis of PGD cycles investigated for aneuploidy from 11/02 to 3/04. MATERIALS AND METHODS: A total of 420 embryos were analyzed for 49 patients. These were from 45 fresh cycles and 10 frozen cycles. Mean age was 37.5 ⫾5.4 and included 24 patients (49%) who were ⱖ40 years old. Embryo morphology were recorded on day 3 and 5. Day 3 embryos, were
Vol. 82, Suppl. 2, September 2004
divided into 3 groups (Grades A: good; B: intermediate; C: poor) based upon standard criteria: number of cells, blastomere size and % cytoplasmic fragmentation. Day 5 embryos were divided into 4 groups (Good blastocyst; Poor blastocyst; Morula; Arrested embryo) based on cell number and morphological quality of the inner cell mass and trophectoderm. Blastomere biopsy was performed on day 3 using standard procedures. FISH using nine probes for chromosomes X, Y, 13, 15, 16, 17, 18, 21 and 22 was applied for aneuploidy screening. Embryo transfer was carried out only if the cycle had normal embryo(s). RESULTS: Chromosomal defects were found in 74% of all Day 3 embryos. There was a trend towards increasing rate of abnormality with poorer morphology (A: 63%; B: 78%; C: 92%). A similar pattern of increased chromosomal abnormalities was also observed with the slower embryo cleavage rates (Table 1). Aneuploidy was found in 73% of the day 5 embryos. Table 2 shows chromosomal abnormality in 42 % of Good blastocysts and 54 % of Poor blastocysts. Twelve cycles (21.8%) out of a total 55 cycles could not be transferred because all of the tested embryos were identified as aneuploid embryos. Mean number of transferred embryos was 1.7 and clinical pregnancy rates in aged (ⱖ 40 years) and younger (⬍ 40 years) patients were 50 % (10/20) and 52 % (12/23) per transfer (or 42% and 48% per cycle), respectively. CONCLUSION: This study suggests embryo morphology correlates only crudely with chromosomal abnormality. Using morphology or cell number failed to separate out a normal embryo population. We conclude that PGD aneuploidy screening may be an alternate way of selecting embryos for transfer. Using aneuploidy screening, we have been able to achieve elevated pregnancy rates in the older patient population. These results support further investigating the use of aneploidy screening for embryo selection. Supported by: None
were then transferred to one of the following culture media: a) Vitro Life Series: IVF500, n⫽4,248; G1.2, n⫽7,323; G1.3, n⫽5,329; or b) Irvine Scientific: P1, n⫽10,805. Embryos were stratified according to growth medium used and patient age (⬍35y and ⱖ35y), and evaluated for development to the ⱖ 8-cell stage. Data were analyzed using Chi square with p⬍0.05 considered statistically significant. RESULTS: Media type significantly impacted development of embryos to ⱖ 8-cell stage (IVF500: 27.7%; G1.2: 34.7%; G1.3: 38.7%; P1: 43.3%; p⬍0.0001). Of note, there was a significant improvement in the % of embryos at ⱖ8-cells as each new generation of Vitro Life medium was introduced (p⬍0.0001). As shown in the table, within age group, medium type continued to impact development to at least the 8-cell stage (p⬍0.0001 for both age groups). However, this impact was not consistently observed within across age groups and within medium.
CONCLUSION: Our data indicate that cleavage rate is influenced by patient age and culture medium type. This relationship should be taken into consideration when establishing quality assurance target values. For example, our current day 5 inclusion criteria were developed using the IVF500 system. Given our current medium system (G1.3) and the results of this analysis, we must now revise our Day 5 inclusion criteria because the relevant target values have changed. Constant surveillance of relevant target values must be maintained in the IVF laboratory since improvements are continually being made in ART systems as demonstrated by the improvement across the generations of Vitro Life media. Supported by: None.
Wednesday, October 20, 2004 5:30 P.M. Wednesday, October 20, 2004 5:15 P.M. O-246 Variables that impact the use of cleavage rates in quality assurance assessment. S. L. Benedict, K. V. Jackson, K. Kelleher, Y. Pan, A. Nureddin, C. Racowsky. Brigham and Women’s Hospital, Harvard Medical School, Boston, MA. OBJECTIVE: Numerous studies have shown that embryo morphology plays a key role in IVF outcome. Development to at least the 8-cell stage has been correlated with improved implantation potential, thereby providing a potentially useful marker for evaluating laboratory performance. However, it is essential to identify what variables impact embryo cleavage when using this type of marker for quality assessment so that relevant target values are identified for each technique being assessed. This study was undertaken to test the hypothesis that culture media type and patient age both influence development of the embryo to at least the 8-cell stage and therefore must be taken into consideration when assessing laboratory performance using ⱖ 8-cell embryos as a marker. DESIGN: Retrospective data analysis. MATERIALS AND METHODS: 7,030 ART cycles performed in our institution from January 1, 1998 to December 31, 2003 were retrospectively reviewed. The study dataset included 3,678 IVF cycles with or without assisted hatching. Oocytes were inseminated in Ham’s F10⫹5% HSA or 10% SSS and evaluated 16 –18 hr later for PN formation. Diploid zygotes
FERTILITY & STERILITY威
O-247 Oolema breakage after ICSI: A technical problem or oocyte quality? I. Aboujaoude, H. Maroun, M. Hamze, F. Hachem, I. Alam, P. Saroufim. Center for Reproductive Medicine and Genetics.Aboujaoude Hospital, Jaleldib, Lebanon. OBJECTIVE: During ICSI the oolema can break and results in oocyte lysis. Oolema breakage after ICSI is underreported and is attributed to a technical difficulty during ICSI procedure or an abnormal pipette diameter. Very few reports have analyzed oolema breakage from oocyte quality. DESIGN: We compared ICSI results (age, length of stimulation, estradiol level, pregnancy rate, miscarriage rate) according to oolema breakage. MATERIALS AND METHODS: 471 couples underwent ICSI cycles after controlled ovarian hyperstimulation. The ICSI procedures were performed by a single technician using Cook® pipette. Group A have less than 15% of oocytes breakage after ICSI when evaluated on day1 and group B have 15% or more oocyte breakage after ICSI. RESULTS: The results are summarized in table 1. In group A smaller number of ampoules was used for ovulation induction than group B. In group A we have less oocytes retrieved and smaller number of immature oocytes. In group A more embryo were obtained than group B. No statistical differences were found between group A and B in terms of implantation rates, pregnancy rates and miscarriage rates. Results table 1
S99