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Efficacy of Cryopreservation of Testicular Sperm in Hypo Spermatogenesis
substitute) at 37°C for 60 minutes. The upper layer containing sperm cells that swim-up from the semen sample was removed for this trial. The swim-up samples was divided into four aliquots. One will be treated with PAF-IUI (Reproductive Biology Associates, Atlanta, GA) preparation at 100nM for 15 minutes at 37 °C and the other with similar medium without PAF. After incubation, sperm cells were assayed for motility characteristics and separated from the PAF-containing or control media by centrifugation through a gradient solution. The washed sperm cell preparations were suspended in 0.5mL of fresh culture medium without PAF. A third aliquot was treated with PAF-IUI continuously and a matching fourth aliquot was exposed only to control medium. All aliquots were assayed at 0, 0.25, 4 and 24 hours for sperm cell motility using computer assisted semen analysis system (Ceros, Hamilton-Thorne, Beverly, MA). Measurements obtained in duplicate of percent progressive motility, track velocity (Vcl), progressive velocity (Vsl), lateral head displacement (ALH), and linearity (LIN) were used to compare treatments with ANOVA. Measurements were obtained by a person blinded to the presence of PAF. RESULTS: Highly motile populations of sperm cells recovered from swim-up preparations of three volunteers were exposure to 100 nM PAF or control solution for 15 minutes and 24 hours. Motility measures including percent progressive motility, Vsl, Vcl, ALH, and LIN did not differ between control and PAF exposure groups. Sperm cell separation following 15 minute exposure to PAF or control medium resulted in recovery of 81% of sperm cells with a reduced (p ⬍ 0.05) motility relative to unmanipulated groups by 24 hours. CONCLUSION: This trial did not demonstrate effects of exposure to 100 nM PAF from a preparation marketed for treatment of human sperm cells prior to insemination. The lack of a detectable effect on an easily assayed sperm parameter, limits the ability to evaluate the quality of this additive to enhance sperm cell function. However, while the absence of measurable effects on simple motility measures does not suggest that PAF is not active in altering other aspects of sperm function, it does mean that more expensive testing of capacitation (such as zona-free sperm penetration or zona binding assays) will be required for laboratory quality control. Supported by: Support provided by Noble Centennial Endowment.
P-841 Regular Coffee Intake is Related to Increased Sperm Motility and Antioxidant Levels in Infertile Men. F. F. Pasqualotto, E. B. Pasqualotto, F. M. Umezu, S. S. Allamaneni, M. Salvador, A. Agarwal. Universidade de Caxias do Sul and Conception, Centro de Reproducao Humana, Caxias do Sul, Brazil; Universidade de Caxias do Sul, Caxias do Sul, Brazil; Cleveland Clinic Foundation, Cleveland, OH. OBJECTIVE: Reactive oxygen species play a significant role in the pathophysiology of male infertility. Superoxide dismutase (SOD) and catalase are important antioxidant enzymes that can quench excess free radicals such as: superoxide anion and hydrogen peroxide, respectively. It is established that sperm motility is higher in patients who regularly drink coffee compared to patients who don’t, however, the mechanism by which caffeine improves sperm motility is not known. The objective of our study was to evaluate and compare the seminal antioxidant enzymatic activity (SOD and catalase levels) among infertile men with the amount of regular coffee intake on a day-to-day basis. DESIGN: Retrospective study at a tertiary care institution. MATERIALS AND METHODS: The Institutional Review Board approved this study. Ten fertile donors and 112 infertile patients were included in the study. Patients were asked about the amount (mL) of regular coffee they drank daily. Semen analysis was performed according to the World Health Organization guidelines and sperm morphology by Tygerberg strict criteria. Superoxide dismutase and catalase levels were determined with a spectrophotometer. RESULTS: A significant difference was noted in the amount of regular coffee drank daily between infertile (200.96 mL ⫾ 46.8) and fertile men (360 mL ⫾ 34.5; P ⫽ 0.043). Significantly lower levels of SOD (14.67 ⫾ 12.27 and 38.03 ⫾ 21.65) and catalase (14.87 ⫾ 16.95 and 34.03 ⫾ 20.65) were seen in infertile patients compared to fertile donors (P ⬍0.0001). A significant correlation between catalase and SOD was observed (r ⫽ 0.461, P ⫽ 0.0001). Sperm morphology by Tygerberg criteria was significantly correlated with the levels of SOD (r ⫽ 0.412, P ⫽ 0.0001) and catalase (r ⫽ 0.315, P ⫽ 0.001). Catalase levels were also correlated with sperm motility
FERTILITY & STERILITY威
(r ⫽ 231, P ⫽ 0.042). Coffee intake was correlated with catalase levels (r ⫽ 0.212, P ⫽ 0.027), but not with SOD levels (r ⫽ 0.173, P ⫽ 0.058). Patients who drank more than 250 mL of coffee on a daily basis had higher catalase levels (39.2 ⫾ 16.2) compared to men who drank less than 250 mL of coffee (17.6 ⫾ 9.2; P ⫽ 0.03). CONCLUSION: The positive correlation between catalase levels, sperm motility and coffee intake on a day-to-day basis may suggest a possible mechanism via caffeine for an increase in sperm motility in men who are coffee drinkers. Supported by: None
P-842 Efficacy of Cryopreservation of Testicular Sperm in Hypo Spermatogenesis. J. Kim, S. Lee, S. Han, S. Song, J. Seo, Y. Park. Laboratory of Reproductive Biology and Infertility, Samsung Cheil Hospital and Women’s Healthcare Center, Seoul, Republic of Korea; Department of Urol. Samsung Cheil Hospital and Women’s Healthcare Center, Sungkyunkwan University School of Medicine, Seoul, Republic of Korea; Laboratory of Reproductive Biology and Infertility, Samsung Cheil Hospital and Women’s Healthcare Center, Sungkyunkwan University School of Medicine, Seoul, Republic of Korea. OBJECTIVE: Testicular sperm using ICSI could achieve optimal fertilization and pregnancy. Hypospermatogenesis is defined as a decrease in the number of spermatogenic cells, so that few mature spermatozoa are formed. This study was performed to assess the fertilizing ability and its embryonic developmental capacity using fresh- and frozen-thawed testicular sperm in hypospermatogenesis patients. DESIGN: Retrospective analysis. MATERIALS AND METHODS: A total eighty four cycle of ICSI was performed with fresh- or frozen-thawed testicular sperm in hypospermatogenesis patients. ICSI with fresh sperm (Group I) was 55 cycles (65.5%) and ICSI with frozen-thawed sperm (Group II) was 29 cycles (34.5%). As we described earlier, testicular tissue was frozen with the programmed cell freezer (Cryomagic I, Miraebiotech, Seoul, Korea). Fertilization check was performed 18⬃20 hrs after ICSI. Embryo grade was assessed on the day 2 and day 3 embryo morphology. Embryo grade was scored six groups and good embryo was classified grade I to grade II. RESULTS: The total fertilization rate with 2-PN was 62.4% and the percentage of cleavage was 95.5%. The development of good embryo was 58.8% (290/493) and pregnancy rate was 39.2% (29/74). In Group I, fertilization rate with 2-PN was 65.7% and the percentage of cleavage was 97.8%. The development of good embryo was 57.6% (204/354) and pregnancy rate was 34.8% (16/46). In Group II, fertilization rate with 2-PN was 54.4% and the percentage of cleavage was 90.3%. The development of good embryo was 61.9% (86/139) and pregnancy rate was 46.4% (13/28). Group II showed higher percentage of good embryo and pregnancy rate than group I. CONCLUSION: In this study we could obtain acceptable fertilization and good embryo rate after ICSI with testicular sperm in hypospermatogenesis. And also frozen sperm showed higher percentage of good embryo and pregnancy rate than fresh sperm. Therefore fresh- and frozen testicular sperm with ICSI is an effective method for embryonic development and pregnancy in azoospermia. Supported by: None
P-843 Forward Progression of Sperm is Predictive of Successful Fertilization With ICSI. M. Traub, S. K. Jindal, C. A. Hickmon, D. Barad, N. Santoro. Albert Einstein College of Medicine, Bronx, NY; Montefiore’s Institute for Reproductive Medicine and Health, Hartsdale, NY. OBJECTIVE: To determine the predictive value of semen parameters for fertilization outcomes in IVF and ICSI cycles. DESIGN: Retrospective study. MATERIALS AND METHODS: This study evaluated 301 initial cycles between January 2002 and December 2003 inclusively. Every patient had a semen analysis followed by either an IVF or ICSI cycle that went to oocytes retrieval. Exclusion criteria were use of donor sperm, TESE cycles, IVF/
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P-841 Regular Coffee Intake is Related to Increased Sperm Motility and Antioxidant Levels in Infertile Men. F. F. Pasqualotto, E. B. Pasqualotto, F. M. Umezu, S. S. Allamaneni, M. Salvador, A. Agarwal. Universidade de Caxias do Sul and Conception, Centro de Reproducao Humana, Caxias do Sul, Brazil; Universidade de Caxias do Sul, Caxias do Sul, Brazil; Cleveland Clinic Foundation, Cleveland, OH. OBJECTIVE: Reactive oxygen species play a significant role in the pathophysiology of male infertility. Superoxide dismutase (SOD) and catalase are important antioxidant enzymes that can quench excess free radicals such as: superoxide anion and hydrogen peroxide, respectively. It is established that sperm motility is higher in patients who regularly drink coffee compared to patients who don’t, however, the mechanism by which caffeine improves sperm motility is not known. The objective of our study was to evaluate and compare the seminal antioxidant enzymatic activity (SOD and catalase levels) among infertile men with the amount of regular coffee intake on a day-to-day basis. DESIGN: Retrospective study at a tertiary care institution. MATERIALS AND METHODS: The Institutional Review Board approved this study. Ten fertile donors and 112 infertile patients were included in the study. Patients were asked about the amount (mL) of regular coffee they drank daily. Semen analysis was performed according to the World Health Organization guidelines and sperm morphology by Tygerberg strict criteria. Superoxide dismutase and catalase levels were determined with a spectrophotometer. RESULTS: A significant difference was noted in the amount of regular coffee drank daily between infertile (200.96 mL ⫾ 46.8) and fertile men (360 mL ⫾ 34.5; P ⫽ 0.043). Significantly lower levels of SOD (14.67 ⫾ 12.27 and 38.03 ⫾ 21.65) and catalase (14.87 ⫾ 16.95 and 34.03 ⫾ 20.65) were seen in infertile patients compared to fertile donors (P ⬍0.0001). A significant correlation between catalase and SOD was observed (r ⫽ 0.461, P ⫽ 0.0001). Sperm morphology by Tygerberg criteria was significantly correlated with the levels of SOD (r ⫽ 0.412, P ⫽ 0.0001) and catalase (r ⫽ 0.315, P ⫽ 0.001). Catalase levels were also correlated with sperm motility
FERTILITY & STERILITY威
(r ⫽ 231, P ⫽ 0.042). Coffee intake was correlated with catalase levels (r ⫽ 0.212, P ⫽ 0.027), but not with SOD levels (r ⫽ 0.173, P ⫽ 0.058). Patients who drank more than 250 mL of coffee on a daily basis had higher catalase levels (39.2 ⫾ 16.2) compared to men who drank less than 250 mL of coffee (17.6 ⫾ 9.2; P ⫽ 0.03). CONCLUSION: The positive correlation between catalase levels, sperm motility and coffee intake on a day-to-day basis may suggest a possible mechanism via caffeine for an increase in sperm motility in men who are coffee drinkers. Supported by: None
P-842 Efficacy of Cryopreservation of Testicular Sperm in Hypo Spermatogenesis. J. Kim, S. Lee, S. Han, S. Song, J. Seo, Y. Park. Laboratory of Reproductive Biology and Infertility, Samsung Cheil Hospital and Women’s Healthcare Center, Seoul, Republic of Korea; Department of Urol. Samsung Cheil Hospital and Women’s Healthcare Center, Sungkyunkwan University School of Medicine, Seoul, Republic of Korea; Laboratory of Reproductive Biology and Infertility, Samsung Cheil Hospital and Women’s Healthcare Center, Sungkyunkwan University School of Medicine, Seoul, Republic of Korea. OBJECTIVE: Testicular sperm using ICSI could achieve optimal fertilization and pregnancy. Hypospermatogenesis is defined as a decrease in the number of spermatogenic cells, so that few mature spermatozoa are formed. This study was performed to assess the fertilizing ability and its embryonic developmental capacity using fresh- and frozen-thawed testicular sperm in hypospermatogenesis patients. DESIGN: Retrospective analysis. MATERIALS AND METHODS: A total eighty four cycle of ICSI was performed with fresh- or frozen-thawed testicular sperm in hypospermatogenesis patients. ICSI with fresh sperm (Group I) was 55 cycles (65.5%) and ICSI with frozen-thawed sperm (Group II) was 29 cycles (34.5%). As we described earlier, testicular tissue was frozen with the programmed cell freezer (Cryomagic I, Miraebiotech, Seoul, Korea). Fertilization check was performed 18⬃20 hrs after ICSI. Embryo grade was assessed on the day 2 and day 3 embryo morphology. Embryo grade was scored six groups and good embryo was classified grade I to grade II. RESULTS: The total fertilization rate with 2-PN was 62.4% and the percentage of cleavage was 95.5%. The development of good embryo was 58.8% (290/493) and pregnancy rate was 39.2% (29/74). In Group I, fertilization rate with 2-PN was 65.7% and the percentage of cleavage was 97.8%. The development of good embryo was 57.6% (204/354) and pregnancy rate was 34.8% (16/46). In Group II, fertilization rate with 2-PN was 54.4% and the percentage of cleavage was 90.3%. The development of good embryo was 61.9% (86/139) and pregnancy rate was 46.4% (13/28). Group II showed higher percentage of good embryo and pregnancy rate than group I. CONCLUSION: In this study we could obtain acceptable fertilization and good embryo rate after ICSI with testicular sperm in hypospermatogenesis. And also frozen sperm showed higher percentage of good embryo and pregnancy rate than fresh sperm. Therefore fresh- and frozen testicular sperm with ICSI is an effective method for embryonic development and pregnancy in azoospermia. Supported by: None
P-843 Forward Progression of Sperm is Predictive of Successful Fertilization With ICSI. M. Traub, S. K. Jindal, C. A. Hickmon, D. Barad, N. Santoro. Albert Einstein College of Medicine, Bronx, NY; Montefiore’s Institute for Reproductive Medicine and Health, Hartsdale, NY. OBJECTIVE: To determine the predictive value of semen parameters for fertilization outcomes in IVF and ICSI cycles. DESIGN: Retrospective study. MATERIALS AND METHODS: This study evaluated 301 initial cycles between January 2002 and December 2003 inclusively. Every patient had a semen analysis followed by either an IVF or ICSI cycle that went to oocytes retrieval. Exclusion criteria were use of donor sperm, TESE cycles, IVF/
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