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Evaluation of silica-based sperm separation method on sperm motility, recovery, and morphology.
G. Pineda, N. T. Serafy. Women’s Research Institute, Wichita, KS; American Assoc of Bioanalysts Proficiency Testing Service, Brownsville, TX. Objective: Determine the level of standardization in the performance of the semen analysis among clinical laboratories in the United States. Design: Anonymous mailed survey. Materials/Methods: A survey was mailed to clinical laboratories enrolled in the American Association of Bioanalysts Andrology Proficiency Testing Program requesting information about the laboratory and the performance of the semen analysis. Responses were received from 536 laboratories. Results: Sixty-one percent of respondents indicated that the laboratory was a part of an assisted reproductive technology program. On average, each laboratory employed 3.4 individuals (range 1–25) to perform semen analyses. The vast majority (94%) of the respondents indicated that their laboratories were accredited. The laboratories performed less than 50 (53%), less than 10 (25%) or less than 5 (16%) andrology laboratory procedures per month. The laboratories routinely report sperm count (94% of laboratories), motility (95%), morphology (85%) and forward progression (69%), and semen volume (96%) as part of the semen analysis. Only 64% of laboratories routinely report abstinence, and 60% of laboratories indicate the criteria used for sperm morphology on the report form. The most common lower limits of normality for sperm count and motility were ⬎20 million sperm/ml (77% of laboratories) and ⬎50% (59%), respectively, although a wide range of “normal values” were reported. Few laboratories performed quality control for sperm counts (29%), motility (28%) and sperm morphology (23%). Approximately one-third (34%) of the laboratories indicated that they were either not familiar with the WHO Laboratory Manual, or did not possess a copy of this manual. Conclusions: These data indicate a significant lack of standardization in the performance and the reporting of semen analyses among laboratories in the United States. Supported by: Women’s Research Institute and the American Association of Bioanalysts.
P-316 Intracytoplasmic sperm injection (ICSI) for everyone: comparison between male factor vs. non-male factor cycles. V. M. Sopelak, S. B. McCulloch, R. S. Hines, B. D. Cowan. Univ of Mississippi Medical Ctr, Jackson, MS. Objective: ICSI is now the gold standard for patients with male factor (MF) infertility. High levels of fertilization “guarantee” embryos for transfer and eliminate the need for second day rescue ICSI when routine insemination leads to fertilization failure. However, the cost versus benefits in patients with semen counts/ml of ⬎20 ⫻ 106 sperm/ml is debatable. The present retrospective study was done to compare and correlate pregnancy outcome and cycle parameters, such as maternal age, oocytes recovered and fertilized and embryos transferred, between MF and non-male factor (nonMF) patients based on sperm count/ml. Design: Retrospective analysis was done on cycles (n ⫽ 227) performed between 1997-2000 where all mature oocytes underwent ICSI (irrespective of diagnosis) and patients had at least 1 embryo transferred. Following analysis of the entire cohort, data were compared between cycles with ⱕ20 ⫻ 106 sperm/ml (MF; n ⫽ 67) versus cycles with ⬎20 ⫻ 106 sperm/ml (non-MF; n ⫽ 160). Materials/Methods: Maternal age, number of oocytes recovered and mature oocytes injected, the number fertilized and embryos transferred and pregnancy outcome were analyzed in these cycles with all ICSI injections. Correlations were performed on the various mentioned parameters before data were regrouped and analyzed by MF versus non-MF. Results: Analysis of the entire 227 cycles showed a significant correlation between maternal age vs number of oocytes recovered (r ⫽ ⫺0.269, P ⬍ 0.001), between the number of oocytes recovered and the number fertilized (r ⫽ 0.762, P ⬍ 0.001) and number of embryos transferred with pregnancy (P ⬍ 0.001). The overall fertilization rate was 76.2%. When data were grouped by MF vs non-MF (see table below), the number of oocytes fertilized, embryos transferred, and pregnancy rates were similar (NS).
S216
Abstracts
Mean ⫾ SEM in ICSI cycles with/without male factor.
Number of cycles Age (years) Oocytes recovered Oocytes injected Number fertilized Embryos transferred Pregnancy rate/cycle
MF
Non-MF
Significance
67 31.4 ⫾ 0.6 11.6 ⫾ 0.9 9.1 ⫾ 0.7 5.6 ⫾ 0.5 2.8 ⫾ 0.1 31.3 ⫾ 5.7%
160 33.2 ⫾ 0.4 8.6 ⫾ 0.4 6.6 ⫾ 0.3 4.8 ⫾ 0.2 2.8 ⫾ 0.1 26.9 ⫾ 3.5%
P ⬍ 0.01 P ⬍ 0.001 P ⬍ 0.001 NS NS NS
Conclusions: In couples with MF infertility, the maternal age of the patients is generally younger yielding more oocytes to inject compared to non-MF patients. Since the number of embryos transferred is restricted to ⱕ3 in patients ⱕ35 years of age, pregnancy rates in both groups are similar. ICSI in non-MF cycles does not increase the pregnancy rate above that found in MF cycles. Supported by: Department of Ob-Gyn.
P-317 Evaluation of silica-based sperm separation method on sperm motility, recovery, and morphology. S. Wang, J. Witmyer, R. Powers. Boston IVF, Waltham, MA. Objective: To evaluate and optimize the efficiency of sperm separation by using a silica-based medium (PureCeption, Sage-Biopharma). Design: Prospective study. Materials/Methods: In previous studies, large variations of sperm parameters (total count, sperm motility, etc) between individuals, even from normozoospermic men, has often lead to large standard deviations (SD) of the parameters measured. The large SD due to individual variability inevitably makes the accurate measurement of outcomes extremely difficult and thus confounds the evaluation of sperm separation methods. To reduce this effect, semen samples of 20 patients were pooled together and divided into equal volume. The advantages of pooling samples are two-fold: To increase the sample volume so more replicates can be performed to reach statistical significance, and more importantly, the individual variability among samples can be minimized. Each aliquot from the pooled sample was loaded into gradients for comparison. Sperm motility, total recovery rate, motile sperm recovery rate, and percentage of normal sperm morphology were measured between (1) two-layer (90%-47%) and three-layer (85.5%-63%45%) PureCeption gradients; (2) large (1.5 ml), medium (0.75 ml), and small (0.3 ml) sizes of three-layer gradients. Results: There was no statistical difference between two-layer and threelayer gradients with respect to motility, total recovery rate, and motile sperm recovery rate. However, three-layer gradients resulted in recovery of a significantly higher percentage of normal sperm morphology compared to two-layer gradients (p ⬍ 0.05). The percentage of normal sperm morphology increased from 2.9% (two-layer gradients) to 5.4% (three-layer gradients) according Tygerberg strict criteria. For different gradient sizes, similar results were found between large, medium, and small gradients in terms of motility and percentage of normal sperm morphology. However, significantly higher sperm recovery rate was observed in the small gradients (14.4%) compared to medium (10.2%) and large (4.9%) columns (p ⬍ 0.05). Conclusions: Sperm samples prepared with three-layer gradients have a higher percentage of normal sperm morphology than samples processed from two-layer gradients. For three-layer gradients, the small size gradient is a better choice since significantly higher sperm recovery rate was obtained, while other parameters like sperm motility and percentage of normal morphology were not compromised.
REPRODUCTIVE SURGERY P-318 Treatment of a unilateral hydrosalpinx by salpingectomy or proximal tubal occlusion may increase the potential for spontaneous pregnancy. A. W. Sagoskin, G. L. Mottla, S. J. Greenhouse, P. E. Browne, R. J.
Vol. 76, No. 3, Suppl. 1, September 2001
P-316 Intracytoplasmic sperm injection (ICSI) for everyone: comparison between male factor vs. non-male factor cycles. V. M. Sopelak, S. B. McCulloch, R. S. Hines, B. D. Cowan. Univ of Mississippi Medical Ctr, Jackson, MS. Objective: ICSI is now the gold standard for patients with male factor (MF) infertility. High levels of fertilization “guarantee” embryos for transfer and eliminate the need for second day rescue ICSI when routine insemination leads to fertilization failure. However, the cost versus benefits in patients with semen counts/ml of ⬎20 ⫻ 106 sperm/ml is debatable. The present retrospective study was done to compare and correlate pregnancy outcome and cycle parameters, such as maternal age, oocytes recovered and fertilized and embryos transferred, between MF and non-male factor (nonMF) patients based on sperm count/ml. Design: Retrospective analysis was done on cycles (n ⫽ 227) performed between 1997-2000 where all mature oocytes underwent ICSI (irrespective of diagnosis) and patients had at least 1 embryo transferred. Following analysis of the entire cohort, data were compared between cycles with ⱕ20 ⫻ 106 sperm/ml (MF; n ⫽ 67) versus cycles with ⬎20 ⫻ 106 sperm/ml (non-MF; n ⫽ 160). Materials/Methods: Maternal age, number of oocytes recovered and mature oocytes injected, the number fertilized and embryos transferred and pregnancy outcome were analyzed in these cycles with all ICSI injections. Correlations were performed on the various mentioned parameters before data were regrouped and analyzed by MF versus non-MF. Results: Analysis of the entire 227 cycles showed a significant correlation between maternal age vs number of oocytes recovered (r ⫽ ⫺0.269, P ⬍ 0.001), between the number of oocytes recovered and the number fertilized (r ⫽ 0.762, P ⬍ 0.001) and number of embryos transferred with pregnancy (P ⬍ 0.001). The overall fertilization rate was 76.2%. When data were grouped by MF vs non-MF (see table below), the number of oocytes fertilized, embryos transferred, and pregnancy rates were similar (NS).
S216
Abstracts
Mean ⫾ SEM in ICSI cycles with/without male factor.
Number of cycles Age (years) Oocytes recovered Oocytes injected Number fertilized Embryos transferred Pregnancy rate/cycle
MF
Non-MF
Significance
67 31.4 ⫾ 0.6 11.6 ⫾ 0.9 9.1 ⫾ 0.7 5.6 ⫾ 0.5 2.8 ⫾ 0.1 31.3 ⫾ 5.7%
160 33.2 ⫾ 0.4 8.6 ⫾ 0.4 6.6 ⫾ 0.3 4.8 ⫾ 0.2 2.8 ⫾ 0.1 26.9 ⫾ 3.5%
P ⬍ 0.01 P ⬍ 0.001 P ⬍ 0.001 NS NS NS
Conclusions: In couples with MF infertility, the maternal age of the patients is generally younger yielding more oocytes to inject compared to non-MF patients. Since the number of embryos transferred is restricted to ⱕ3 in patients ⱕ35 years of age, pregnancy rates in both groups are similar. ICSI in non-MF cycles does not increase the pregnancy rate above that found in MF cycles. Supported by: Department of Ob-Gyn.
P-317 Evaluation of silica-based sperm separation method on sperm motility, recovery, and morphology. S. Wang, J. Witmyer, R. Powers. Boston IVF, Waltham, MA. Objective: To evaluate and optimize the efficiency of sperm separation by using a silica-based medium (PureCeption, Sage-Biopharma). Design: Prospective study. Materials/Methods: In previous studies, large variations of sperm parameters (total count, sperm motility, etc) between individuals, even from normozoospermic men, has often lead to large standard deviations (SD) of the parameters measured. The large SD due to individual variability inevitably makes the accurate measurement of outcomes extremely difficult and thus confounds the evaluation of sperm separation methods. To reduce this effect, semen samples of 20 patients were pooled together and divided into equal volume. The advantages of pooling samples are two-fold: To increase the sample volume so more replicates can be performed to reach statistical significance, and more importantly, the individual variability among samples can be minimized. Each aliquot from the pooled sample was loaded into gradients for comparison. Sperm motility, total recovery rate, motile sperm recovery rate, and percentage of normal sperm morphology were measured between (1) two-layer (90%-47%) and three-layer (85.5%-63%45%) PureCeption gradients; (2) large (1.5 ml), medium (0.75 ml), and small (0.3 ml) sizes of three-layer gradients. Results: There was no statistical difference between two-layer and threelayer gradients with respect to motility, total recovery rate, and motile sperm recovery rate. However, three-layer gradients resulted in recovery of a significantly higher percentage of normal sperm morphology compared to two-layer gradients (p ⬍ 0.05). The percentage of normal sperm morphology increased from 2.9% (two-layer gradients) to 5.4% (three-layer gradients) according Tygerberg strict criteria. For different gradient sizes, similar results were found between large, medium, and small gradients in terms of motility and percentage of normal sperm morphology. However, significantly higher sperm recovery rate was observed in the small gradients (14.4%) compared to medium (10.2%) and large (4.9%) columns (p ⬍ 0.05). Conclusions: Sperm samples prepared with three-layer gradients have a higher percentage of normal sperm morphology than samples processed from two-layer gradients. For three-layer gradients, the small size gradient is a better choice since significantly higher sperm recovery rate was obtained, while other parameters like sperm motility and percentage of normal morphology were not compromised.
REPRODUCTIVE SURGERY P-318 Treatment of a unilateral hydrosalpinx by salpingectomy or proximal tubal occlusion may increase the potential for spontaneous pregnancy. A. W. Sagoskin, G. L. Mottla, S. J. Greenhouse, P. E. Browne, R. J.
Vol. 76, No. 3, Suppl. 1, September 2001