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Enzymatic digestion of tissue in azoospermic, non-obstructed men undergoing testicular sperm extraction (TESE) for in vitro fertilization.
implantation rates were 13.7% and 19.7% (both NS) in the two groups respectively (biopsy and control groups). Conclusions: A statistically not significantly lower pregnancy and implantation rates were observed in the group of patients where embryo biopsy procedure was performed and a single blastomere was removed. An insufficient number of patients enrolled into the study and/or disadvantageous side effects of the biopsy procedure may explain this difference. This might be clarified better in the near future by continuing the present study including a larger number of eligible patients. In conclusion, in order to increase implantation rates in older patients by means of PGD of aneuploidy, the advantage of selecting normal embryos after biopsy for transfer should compensate in excess for the possible damage caused by the blastomere removal procedure. Preliminary results from another study suggest that this occurs in patients 37 and older where the rate of aneuploidy starts to be considerable.
ART: MALE FACTOR P-74 Fine needle testicular sperm aspiration: an alternative to open testicular biopsy in patients with non-obstructive azoospermia. Z. A. Nassar, D. Lakkis, M. Sasy, I. Khan, M. Abuzeid, M. H. Fakih. IVF Michigan, Rochester Hills, MI. Objective: Open testicular biopsy is still the most commonly performed method for surgical sperm retrieval in patients with non-obstructive azoospermia. The aim of this study is to compare the reproductive outcome of intracytoplasmic sperm injection (ICSI) cycles following surgical sperm retrieval with testicular aspiration versus open testicular biopsy. Design: Retrospective study. Materials/Methods: Between May 1994 and December 2000, a total of 96 female patients whose partner had non-obstructive azoospermia underwent 120 cycles of ICSI following surgical sperm retrieval. Only women 35 years old or less and whose partner had a previous diagnostic testicular biopsy were included (n ⫽ 60 patients, 75 cycles). They were divided into 2 groups based on the technique of sperm retrieval. Group 1 (n ⫽ 49 cycles) included those whose partner underwent fine needle testicular sperm aspiration and Group 2 (n ⫽ 26 cycles) included those who underwent open testicular biopsy. Mean female age was 30.3 ⫾ 4.8 vs 29.4 ⫾ 4.0 respectively and mean male age was 35.9 ⫾ 5.0 vs 36.6 ⫾ 4.9 respectively. Hypospermatogenesis was diagnosed in 25 (51.0%) cycles in group 1 vs 14 (53.8%) cycles in group 2, germ cell aplasia in 10 (20.4%) vs 6 (23.1%), incomplete maturation arrest in 8 (16.3%) vs 5 (19.2%), complete arrest in 5 (10.2%) vs 1 (3.8%) respectively and setoli-cell only syndrome in 1 (2.0%) cycle in group 1. Results: There was no statistically significant difference in female age, male age, previous histopathology, downregulation and stimulation protocol, number of eggs retrieved, number and quality of embryos transferred, and technique of embryo transfer (ET) between the 2 groups. Outcome of ICSI following surgical sperm retrieval in the 2 groups is shown in Table 1. There was no statistically significant difference in sperm recovery rate, fertilization rate, implantation rate, and pregnancy rate between the 2 groups. Immature sperm was retrieved and injected in 20 (40.8%) cycles in group 1 compared to 11 (42.3%) in group 2 and no sperm could be retrieved in 9 (18.4%) cycles in group 1 vs 4 (15.4%) in group 2.The difference was not statistically significant. Open biopsy performed at the same setting in the 9 cycles in group 1 who failed sperm aspiration was not successful in mature sperm retrieval as well. Table 1. Outcome of ICSI following surgical sperm retrieval with aspiration vs biopsy in non-obstructive azoospermia.
Motile sperm recovery rate Fertilization rate Implantation rate Clinical pregnancy rate per cycle Clinical pregnancy rate per ET Delivery/ongoing pregnancy rate per cycle Delivery/ongoing pregnancy rate per ET
FERTILITY & STERILITY威
Group 1: Aspiration (n⫽49)
Group 2: biopsy (n ⫽ 26)
24.5 36.4 8.2 14.3 20.0 8.2 11.4
19.2 31.3 5.6 11.5 17.6 11.5 17.6
Conclusions: Our data suggests that fine needle testicular sperm aspiration could be an acceptable alternative to open testicular biopsy in patients with non-obstructive azoospermia with different testicular histopathology.
P-75 Enzymatic digestion of tissue in azoospermic, non-obstructed men undergoing testicular sperm extraction (TESE) for in vitro fertilization. C. Cook, D. Liotta, P. Schlegel, Z. Ye, L. L. Veeck. Weill Medical Coll of Cornell Univ, New York, NY. Objective: To examine the efficiency of enzymatic treatment of testicular tissue as a tool for isolating adequate numbers of viable spermatozoa for intracytoplasmic sperm injection (ICSI). Design: Retrospective analysis of in vitro fertilization (IVF) cycles where spermatozoa were difficult to isolate from testicular tissue, requiring more aggressive procedures for identification and isolation. Materials/Methods: Between March 1999 and December 2000, testicular biopsy was carried out on 142 non-obstructed men for IVF treatment. Of these, the tissue from 57 men was subjected to digestion techniques in an effort to increase the opportunity for sperm recovery. In 41 patients with tissue digestion, TESE was performed the same day as oocyte harvest in the mens’ partners; the remaining 16 patients had TESE performed one or two days in advance. When no sperm were initially identified after thorough mechanical maceration of specimens in the operating room and sedimentation of tissue fractions, tissue digestion was carried out using 2.6 mg/mL collagenase (Sigma Chemical Company; #C1889) and 25 g/mL DNAse (Sigma; #DN-25). All couples were offered the option of using donor sperm as a backup measure in the event of failing to find sperm after tissue digestion. Results: In 16 cases where rare spermatozoa were identified after sedimentation procedures, all resulted in enhanced recovery of additional sperm after treatment of tissue with collagenase and DNAse. Nonetheless, normal fertilization was only 42% in this group and clinical pregnancy after transfer was a disappointing 36%. In 41 cases where spermatozoa were not found in the initial workup, tissue digestion did not enhance sperm recovery efforts. Twenty of these couples opted for donor sperm after TESE failure, resulting in a fertilization rate of 71% and a clinical pregnancy rate of 61%. Nine TESE failures refused donor sperm as back-up, and because TESE was performed before oocytes were collected, it was possible to cancel oocyte harvest. Twelve couples had oocytes harvested needlessly from the wife because of suboptimal timing of the TESE procedure. These 12 patients, who did not accept the option of using donor semen, had no sperm for injection and thus, oocytes were simply discarded. Conclusions: Enzymatic digestion of testicular tissue may aid in isolating viable spermatozoa for the extremely difficult TESE case with rare spermatozoa, but is not an effective treatment for non-obstructive, azoospermic men if no spermatozoa have been identified after initial sedimentation and searching. Because of disappointing outcomes with prolonged search and digestion efforts when initial sperm are not identified, couples with poor prognosis for sperm recovery should have TESE procedures performed at least several hours, and up to one day in advance of surgical procedures for oocyte harvest. Supported by: The Center for Reproductive Medicine and Infertility.
P-76 Variations of seminal characteristics in young and elderly men: Is the aging effect real? P. M. Zavos, K. Kaskar, J. R. Correa, P. N. Zarmakoupis-Zavos. The Kentucky Ctr for Reproductive Medicine and IVF and the Andrology Institute of America, Lexington, KY; Ctr de Fertilidad del Caribe, Rio Piedras, Puerto Rico. Objective: It has been shown very clearly by now that aging in women significantly reduces the potential to produce high quality oocytes and achieve conception. In men, the results of aging on spermatogenesis and ejaculate characteristics have been somewhat mixed. Meanwhile, reproductive ability is decreased in aging animals. The effects of age in bulls were greatest for sperm concentration per mL and total sperm output. We have previously shown that there is a decrease in the total motile sperm (TMS) and total functional sperm fraction (TFSF) with increasing age, indicating an aging effect on seminal characteristics of normospermic men. The aim of
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ART: MALE FACTOR P-74 Fine needle testicular sperm aspiration: an alternative to open testicular biopsy in patients with non-obstructive azoospermia. Z. A. Nassar, D. Lakkis, M. Sasy, I. Khan, M. Abuzeid, M. H. Fakih. IVF Michigan, Rochester Hills, MI. Objective: Open testicular biopsy is still the most commonly performed method for surgical sperm retrieval in patients with non-obstructive azoospermia. The aim of this study is to compare the reproductive outcome of intracytoplasmic sperm injection (ICSI) cycles following surgical sperm retrieval with testicular aspiration versus open testicular biopsy. Design: Retrospective study. Materials/Methods: Between May 1994 and December 2000, a total of 96 female patients whose partner had non-obstructive azoospermia underwent 120 cycles of ICSI following surgical sperm retrieval. Only women 35 years old or less and whose partner had a previous diagnostic testicular biopsy were included (n ⫽ 60 patients, 75 cycles). They were divided into 2 groups based on the technique of sperm retrieval. Group 1 (n ⫽ 49 cycles) included those whose partner underwent fine needle testicular sperm aspiration and Group 2 (n ⫽ 26 cycles) included those who underwent open testicular biopsy. Mean female age was 30.3 ⫾ 4.8 vs 29.4 ⫾ 4.0 respectively and mean male age was 35.9 ⫾ 5.0 vs 36.6 ⫾ 4.9 respectively. Hypospermatogenesis was diagnosed in 25 (51.0%) cycles in group 1 vs 14 (53.8%) cycles in group 2, germ cell aplasia in 10 (20.4%) vs 6 (23.1%), incomplete maturation arrest in 8 (16.3%) vs 5 (19.2%), complete arrest in 5 (10.2%) vs 1 (3.8%) respectively and setoli-cell only syndrome in 1 (2.0%) cycle in group 1. Results: There was no statistically significant difference in female age, male age, previous histopathology, downregulation and stimulation protocol, number of eggs retrieved, number and quality of embryos transferred, and technique of embryo transfer (ET) between the 2 groups. Outcome of ICSI following surgical sperm retrieval in the 2 groups is shown in Table 1. There was no statistically significant difference in sperm recovery rate, fertilization rate, implantation rate, and pregnancy rate between the 2 groups. Immature sperm was retrieved and injected in 20 (40.8%) cycles in group 1 compared to 11 (42.3%) in group 2 and no sperm could be retrieved in 9 (18.4%) cycles in group 1 vs 4 (15.4%) in group 2.The difference was not statistically significant. Open biopsy performed at the same setting in the 9 cycles in group 1 who failed sperm aspiration was not successful in mature sperm retrieval as well. Table 1. Outcome of ICSI following surgical sperm retrieval with aspiration vs biopsy in non-obstructive azoospermia.
Motile sperm recovery rate Fertilization rate Implantation rate Clinical pregnancy rate per cycle Clinical pregnancy rate per ET Delivery/ongoing pregnancy rate per cycle Delivery/ongoing pregnancy rate per ET
FERTILITY & STERILITY威
Group 1: Aspiration (n⫽49)
Group 2: biopsy (n ⫽ 26)
24.5 36.4 8.2 14.3 20.0 8.2 11.4
19.2 31.3 5.6 11.5 17.6 11.5 17.6
Conclusions: Our data suggests that fine needle testicular sperm aspiration could be an acceptable alternative to open testicular biopsy in patients with non-obstructive azoospermia with different testicular histopathology.
P-75 Enzymatic digestion of tissue in azoospermic, non-obstructed men undergoing testicular sperm extraction (TESE) for in vitro fertilization. C. Cook, D. Liotta, P. Schlegel, Z. Ye, L. L. Veeck. Weill Medical Coll of Cornell Univ, New York, NY. Objective: To examine the efficiency of enzymatic treatment of testicular tissue as a tool for isolating adequate numbers of viable spermatozoa for intracytoplasmic sperm injection (ICSI). Design: Retrospective analysis of in vitro fertilization (IVF) cycles where spermatozoa were difficult to isolate from testicular tissue, requiring more aggressive procedures for identification and isolation. Materials/Methods: Between March 1999 and December 2000, testicular biopsy was carried out on 142 non-obstructed men for IVF treatment. Of these, the tissue from 57 men was subjected to digestion techniques in an effort to increase the opportunity for sperm recovery. In 41 patients with tissue digestion, TESE was performed the same day as oocyte harvest in the mens’ partners; the remaining 16 patients had TESE performed one or two days in advance. When no sperm were initially identified after thorough mechanical maceration of specimens in the operating room and sedimentation of tissue fractions, tissue digestion was carried out using 2.6 mg/mL collagenase (Sigma Chemical Company; #C1889) and 25 g/mL DNAse (Sigma; #DN-25). All couples were offered the option of using donor sperm as a backup measure in the event of failing to find sperm after tissue digestion. Results: In 16 cases where rare spermatozoa were identified after sedimentation procedures, all resulted in enhanced recovery of additional sperm after treatment of tissue with collagenase and DNAse. Nonetheless, normal fertilization was only 42% in this group and clinical pregnancy after transfer was a disappointing 36%. In 41 cases where spermatozoa were not found in the initial workup, tissue digestion did not enhance sperm recovery efforts. Twenty of these couples opted for donor sperm after TESE failure, resulting in a fertilization rate of 71% and a clinical pregnancy rate of 61%. Nine TESE failures refused donor sperm as back-up, and because TESE was performed before oocytes were collected, it was possible to cancel oocyte harvest. Twelve couples had oocytes harvested needlessly from the wife because of suboptimal timing of the TESE procedure. These 12 patients, who did not accept the option of using donor semen, had no sperm for injection and thus, oocytes were simply discarded. Conclusions: Enzymatic digestion of testicular tissue may aid in isolating viable spermatozoa for the extremely difficult TESE case with rare spermatozoa, but is not an effective treatment for non-obstructive, azoospermic men if no spermatozoa have been identified after initial sedimentation and searching. Because of disappointing outcomes with prolonged search and digestion efforts when initial sperm are not identified, couples with poor prognosis for sperm recovery should have TESE procedures performed at least several hours, and up to one day in advance of surgical procedures for oocyte harvest. Supported by: The Center for Reproductive Medicine and Infertility.
P-76 Variations of seminal characteristics in young and elderly men: Is the aging effect real? P. M. Zavos, K. Kaskar, J. R. Correa, P. N. Zarmakoupis-Zavos. The Kentucky Ctr for Reproductive Medicine and IVF and the Andrology Institute of America, Lexington, KY; Ctr de Fertilidad del Caribe, Rio Piedras, Puerto Rico. Objective: It has been shown very clearly by now that aging in women significantly reduces the potential to produce high quality oocytes and achieve conception. In men, the results of aging on spermatogenesis and ejaculate characteristics have been somewhat mixed. Meanwhile, reproductive ability is decreased in aging animals. The effects of age in bulls were greatest for sperm concentration per mL and total sperm output. We have previously shown that there is a decrease in the total motile sperm (TMS) and total functional sperm fraction (TFSF) with increasing age, indicating an aging effect on seminal characteristics of normospermic men. The aim of
S137