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Varicocelectomy for infertile couples with advanced paternal age
values for disappearance of gynecomastia were 13.0 0.86, 164.5 5.6, 48.62 5.8, 12.5 4.2, 13.5 4.0 respectively. In 11 year olds, when gynecomastia was seen for the first time, it was positively correlated with varicocele (odds ratio ¼ 3.039, 95% confidence interval ¼ 1.215 – 7.598). However no significant correlation was seen in subsequent years which may be due to the transient nature of gynecomastia. CONCLUSIONS: Adolescent gynecomastia is a transient mid-puberty event. Gynecomastia is more likely to develop in young boys having varicocele. Supported by: None.
Tuesday, October 16, 2007 3:45 pm O-131 PROLIFERATION AND DIFFERENTIATION OF MALE GERMLINE STEM CELLS FROM TESTES OF NON-OBSTRUCTIVE AZOOSPERMIA USING SEQUENTIAL CULTURE SYSTEMS. J. J. Lim, D. R. Lee, H. J. Kim, S. H. Song, T. K. Yoon, K. S. Kim. Fertility Center of CHA General Hospital, CHA Research Institute, Pochon CHA University, Seoul, Korea; Department of Anatomy and Cell Biology, College of Medicine, Hanyang University, Seoul, Korea. OBJECTIVE: Male germ-line stem cells (GSCs, spermatogonial stem cells) self-renew and produce large numbers of differentiating germ cells that become spermatozoa throughout the adult life. In humans, our group reported that GSCs had a potential to conceive of non-obstructive azoospermia, but no pregnancy may be resulted from low number of isolated GSCs and in vitro differentiated haploid germ cells (Lee et al., 2006). DESIGN: This study was to develop the culture methods for enrichment of male GSCs and differentiating into sperm lineage cells. MATERIALS AND METHODS: Testicular tissues were obtained from 35 non-obstructive azoosperimic patients (maturation arrest and Sertoli cellonly syndrome) under informed consent were dissociated and plated into gelatin-coated dishes using sequentially different culture medium: 1) GSC-C medium for GSC colony formation (DMEM containing leukemia inhibitor factor, bFGF, and 10% fetal bovine serum; Lee et al., 2006), 2) GSC-P medium for GSC proliferation (StemPro-34TM containing GDNF, EGF, bFGF, and LIF; Kanatsu-Shinohara et al., 2003), 3) GSC-D system for differentiation (DMEM/F12 containing testosterone, vitamins, and 10% bovine calf serum; Lee et al., 2001) and calcium alginate encapsulation. RESULTS: During cultivation in GSC-C medium, largely (>200 mm) and tightly attached multi-cellular colonies were formed from testicular cells of 10 patients (28.6%) at first passage during 2–3 weeks. From 2nd passages using GSC-P medium, partially dissociated colonies (<80 mm) were collected and then passaged every 2 weeks. After passaging, cell clumps were analyzed by RT-PCR, immunocytochemistry, flow cytometry using markers of GSCs (integrin a6, and ß1) and post-meiotic germ cell (c-Kit receptor). From 2nd passages, the positive cells for integrin a6/ß1 were gradually increased until 5 passages. Expression of c-kit receptors were still limited by 5th passages. During cultivation in GSC-D system, c-kit positive cells that represent post-meiotic germ cell were dramatically increased compared to the cells cultured in GSC-P medium. CONCLUSIONS: GSCs from testicular cells of non-obstructive azoospermic patients were proliferated and could be differentiated into sperm lineage cells in our sequentially different culture media. Therefore, our sequential culture system may provide in vitro model system for treatment of male infertility. Supported by: The National R&D Program (2006–04127) of the Ministry of Science & Technology, Republic of Korea and the Korea Science and Engineering Foundation.
Tuesday, October 16, 2007 4:00 pm O-132 COMBINED EFFECTS OF THE EXPERIMENTAL LEFT VARICOCELE AND LEAD OR NICOTINE ON THE RAT TESTIS. B. R. Emery, Y. Sun, D. T. Carrell. Andrology And IVF Laboratories, Univ. of Utah School of Medicine, Salt Lake City, UT; ObGyn, Division of Reproductive Endocrinology, Univ. of Utah School of Medicine, Salt Lake City, UT.
S50
Abstracts
OBJECTIVE: The purpose of this study was to evaluate if the toxic effects of lead or nicotine on the testis are exacerbated by the presence of a left-sided varicocele and are they intensified over a period of chronic exposure. DESIGN: This animal study used the experimental left varicocele (ELV) model in the rat over a period of nine months. Animals were divided into groups of control animals (no treatment), sham operated (exposure of the renal vein/spermatic vein junction), and experimentally induced left varicocele (partial ligation of the left spermatic vein as it empties into the left renal vein). These three groups were then divided into animals receiving lead, nicotine, or no drug. The animals were then followed for three, six or nine months at which time the testicular status was evaluated. MATERIALS AND METHODS: The left testicular varicocele was induced following the procedure of Turner, TT (2001). Specified rats were treated with lead acetate at 0.6% in distilled water ad libitum QD or ip injection of nicotine at 0.25 mg/100 g QOD. Rats were then chosen at intervals of three months for collection of the testis and epididymis from the left and right side. Testis and body weights were made; the epididymis was weighed, minced, and incubated in Quinn’s Sperm Wash Media for 30 minutes. The sperm motility and concentration were noted. Measured parameters were evaluated for statistical significance between groups using STATA 9.0 for Macintosh. RESULTS: Control rats (n ¼ 18) showed a statistically significant difference in the outcomes from the treated rats but not sham-operated animals. The treatment groups consisted of 84 rats with ELV; eight rats failed the ELV or had complications. The remaining 90.4% of the animals had an increased dilation of the left spermatic vein of 1.0–2.0 mm from a baseline diameter of 0.3 mm. The rats in the treatment groups showed a decrease in testicular weight, which showed a consistant trend over the period of study. The concentration of sperm recovered from the caudal epididymis was correlated to the testicular weight and was reduced in treated animals. CONCLUSIONS: These data support the hypothesis that the left varicocele does alter testicular efficiency and may cause an increased risk to those men who are also exposed to testicular toxicants from environmental contaminants or lifestyle choices. Supported by: None.
Tuesday, October 16, 2007 4:15 pm O-133 VARICOCELECTOMY FOR INFERTILE COUPLES WITH ADVANCED PATERNAL AGE. A. Grunbaum, J. Boman, A. Zini. Urology, McGill University, Montreal, QC, Canada. OBJECTIVE: Varicocele represents one of the most common causes of correctable male factor infertility. Most studies indicate that varicocelectomy has a positive effect on male fertility potential and pregnancy outcome. However, there is limited data on the impact of paternal age on semen parameters and pregnancy outcomes after varicocelectomy. We sought to evaluate the clinical outcomes of infertile couples with a clinical varicocele and advanced paternal age. DESIGN: Retrospective review. MATERIALS AND METHODS: We performed a retrospective review of 581 consecutive infertile couples in whom the male partner presented with a clinical varicocele and an abnormal semen analysis. We compared the clinical characteristics and treatment outcomes of older (40 years of age and older, n ¼ 115) to younger men (under 40, n ¼ 466). 60 older men and 295 younger men underwent subinguinal microsurgical varicocelectomy. RESULTS: The proportion of men with secondary infertility was significantly higher in the men R40 compared to the men <40 (44% [51/115] vs. 19% [87/466], respectively, P<0.001). On the other hand, the proportion of men who elected to undergo surgery was significantly lower in the men R40 compared to the men <40 (52% [60/115] vs. 63% [295/466], respectively, P<0.05). There were no significant differences in baseline semen and clinical parameters and in pregnancy rates after varicocelectomy in men R40 compared to men <40. However, the spontaneous pregnancy rate in men R40 who underwent varicocelectomy was significantly greater than that of an age-matched control group who did not undergo surgery (50% vs. 21%, respectively, P<0.05). CONCLUSIONS: This study on the natural history of infertile men with varicocele suggests that paternal age does not adversely impact the response to varicocelectomy. The data support the practice of varicocelectomy for treatment of clinical varicocele in men over the age of 40 with the intent of improving pregnancy rates. Supported by: None.
Vol. 88, Suppl 1, September 2007
Tuesday, October 16, 2007 3:45 pm O-131 PROLIFERATION AND DIFFERENTIATION OF MALE GERMLINE STEM CELLS FROM TESTES OF NON-OBSTRUCTIVE AZOOSPERMIA USING SEQUENTIAL CULTURE SYSTEMS. J. J. Lim, D. R. Lee, H. J. Kim, S. H. Song, T. K. Yoon, K. S. Kim. Fertility Center of CHA General Hospital, CHA Research Institute, Pochon CHA University, Seoul, Korea; Department of Anatomy and Cell Biology, College of Medicine, Hanyang University, Seoul, Korea. OBJECTIVE: Male germ-line stem cells (GSCs, spermatogonial stem cells) self-renew and produce large numbers of differentiating germ cells that become spermatozoa throughout the adult life. In humans, our group reported that GSCs had a potential to conceive of non-obstructive azoospermia, but no pregnancy may be resulted from low number of isolated GSCs and in vitro differentiated haploid germ cells (Lee et al., 2006). DESIGN: This study was to develop the culture methods for enrichment of male GSCs and differentiating into sperm lineage cells. MATERIALS AND METHODS: Testicular tissues were obtained from 35 non-obstructive azoosperimic patients (maturation arrest and Sertoli cellonly syndrome) under informed consent were dissociated and plated into gelatin-coated dishes using sequentially different culture medium: 1) GSC-C medium for GSC colony formation (DMEM containing leukemia inhibitor factor, bFGF, and 10% fetal bovine serum; Lee et al., 2006), 2) GSC-P medium for GSC proliferation (StemPro-34TM containing GDNF, EGF, bFGF, and LIF; Kanatsu-Shinohara et al., 2003), 3) GSC-D system for differentiation (DMEM/F12 containing testosterone, vitamins, and 10% bovine calf serum; Lee et al., 2001) and calcium alginate encapsulation. RESULTS: During cultivation in GSC-C medium, largely (>200 mm) and tightly attached multi-cellular colonies were formed from testicular cells of 10 patients (28.6%) at first passage during 2–3 weeks. From 2nd passages using GSC-P medium, partially dissociated colonies (<80 mm) were collected and then passaged every 2 weeks. After passaging, cell clumps were analyzed by RT-PCR, immunocytochemistry, flow cytometry using markers of GSCs (integrin a6, and ß1) and post-meiotic germ cell (c-Kit receptor). From 2nd passages, the positive cells for integrin a6/ß1 were gradually increased until 5 passages. Expression of c-kit receptors were still limited by 5th passages. During cultivation in GSC-D system, c-kit positive cells that represent post-meiotic germ cell were dramatically increased compared to the cells cultured in GSC-P medium. CONCLUSIONS: GSCs from testicular cells of non-obstructive azoospermic patients were proliferated and could be differentiated into sperm lineage cells in our sequentially different culture media. Therefore, our sequential culture system may provide in vitro model system for treatment of male infertility. Supported by: The National R&D Program (2006–04127) of the Ministry of Science & Technology, Republic of Korea and the Korea Science and Engineering Foundation.
Tuesday, October 16, 2007 4:00 pm O-132 COMBINED EFFECTS OF THE EXPERIMENTAL LEFT VARICOCELE AND LEAD OR NICOTINE ON THE RAT TESTIS. B. R. Emery, Y. Sun, D. T. Carrell. Andrology And IVF Laboratories, Univ. of Utah School of Medicine, Salt Lake City, UT; ObGyn, Division of Reproductive Endocrinology, Univ. of Utah School of Medicine, Salt Lake City, UT.
S50
Abstracts
OBJECTIVE: The purpose of this study was to evaluate if the toxic effects of lead or nicotine on the testis are exacerbated by the presence of a left-sided varicocele and are they intensified over a period of chronic exposure. DESIGN: This animal study used the experimental left varicocele (ELV) model in the rat over a period of nine months. Animals were divided into groups of control animals (no treatment), sham operated (exposure of the renal vein/spermatic vein junction), and experimentally induced left varicocele (partial ligation of the left spermatic vein as it empties into the left renal vein). These three groups were then divided into animals receiving lead, nicotine, or no drug. The animals were then followed for three, six or nine months at which time the testicular status was evaluated. MATERIALS AND METHODS: The left testicular varicocele was induced following the procedure of Turner, TT (2001). Specified rats were treated with lead acetate at 0.6% in distilled water ad libitum QD or ip injection of nicotine at 0.25 mg/100 g QOD. Rats were then chosen at intervals of three months for collection of the testis and epididymis from the left and right side. Testis and body weights were made; the epididymis was weighed, minced, and incubated in Quinn’s Sperm Wash Media for 30 minutes. The sperm motility and concentration were noted. Measured parameters were evaluated for statistical significance between groups using STATA 9.0 for Macintosh. RESULTS: Control rats (n ¼ 18) showed a statistically significant difference in the outcomes from the treated rats but not sham-operated animals. The treatment groups consisted of 84 rats with ELV; eight rats failed the ELV or had complications. The remaining 90.4% of the animals had an increased dilation of the left spermatic vein of 1.0–2.0 mm from a baseline diameter of 0.3 mm. The rats in the treatment groups showed a decrease in testicular weight, which showed a consistant trend over the period of study. The concentration of sperm recovered from the caudal epididymis was correlated to the testicular weight and was reduced in treated animals. CONCLUSIONS: These data support the hypothesis that the left varicocele does alter testicular efficiency and may cause an increased risk to those men who are also exposed to testicular toxicants from environmental contaminants or lifestyle choices. Supported by: None.
Tuesday, October 16, 2007 4:15 pm O-133 VARICOCELECTOMY FOR INFERTILE COUPLES WITH ADVANCED PATERNAL AGE. A. Grunbaum, J. Boman, A. Zini. Urology, McGill University, Montreal, QC, Canada. OBJECTIVE: Varicocele represents one of the most common causes of correctable male factor infertility. Most studies indicate that varicocelectomy has a positive effect on male fertility potential and pregnancy outcome. However, there is limited data on the impact of paternal age on semen parameters and pregnancy outcomes after varicocelectomy. We sought to evaluate the clinical outcomes of infertile couples with a clinical varicocele and advanced paternal age. DESIGN: Retrospective review. MATERIALS AND METHODS: We performed a retrospective review of 581 consecutive infertile couples in whom the male partner presented with a clinical varicocele and an abnormal semen analysis. We compared the clinical characteristics and treatment outcomes of older (40 years of age and older, n ¼ 115) to younger men (under 40, n ¼ 466). 60 older men and 295 younger men underwent subinguinal microsurgical varicocelectomy. RESULTS: The proportion of men with secondary infertility was significantly higher in the men R40 compared to the men <40 (44% [51/115] vs. 19% [87/466], respectively, P<0.001). On the other hand, the proportion of men who elected to undergo surgery was significantly lower in the men R40 compared to the men <40 (52% [60/115] vs. 63% [295/466], respectively, P<0.05). There were no significant differences in baseline semen and clinical parameters and in pregnancy rates after varicocelectomy in men R40 compared to men <40. However, the spontaneous pregnancy rate in men R40 who underwent varicocelectomy was significantly greater than that of an age-matched control group who did not undergo surgery (50% vs. 21%, respectively, P<0.05). CONCLUSIONS: This study on the natural history of infertile men with varicocele suggests that paternal age does not adversely impact the response to varicocelectomy. The data support the practice of varicocelectomy for treatment of clinical varicocele in men over the age of 40 with the intent of improving pregnancy rates. Supported by: None.
Vol. 88, Suppl 1, September 2007