- Email: [email protected]
Tese-ICSI in 62 postchemotherapy persistent azoospermic men
MALE FACTOR P-970 Thursday, October 17, 2013 TESE-ICSI IN 62 POSTCHEMOTHERAPY PERSISTENT AZOOSPERMIC MEN. H. Okada, T. Iwahata, K. Suzuki, S. Ohta, T. Shin, Y. Kobori. Urology, Dokkyo Medical University Koshigaya Hospital, Koshigaya, Saitama, Japan. OBJECTIVE: Persistent azoospermic patients after anti-cancer chemotherapy have been considered sterile. We report our experience with testicular sperm extraction (TESE) combined with intracytoplasmic sperm injection (ICSI) applied to postchemotherapy azoospermic men. DESIGN: Retrospective chart review and interview through telephone. MATERIALS AND METHODS: Between 1999 and 2011 a total of 62 patients who previously received chemotherapy for malignant diseases underwent TESE. TESE procedures were performed with microdissection TESE by a single surgeon under general or spinal anesthesia. The hormonal profile of the first visit to the male infertility clinic, histology of testicular biopsies, and outcomes of TESE-ICSI in this subgroup of patients were analyzed. RESULTS: Twenty-one patients had received chemotherapy for testicular canner, seven patients for non Hodgkin lymphoma, nine patients for Hodgkin lymphoma, nine patients for acute lymphoblastic leukemia, six patients for acute myeloblastic leukemia, seven patients for rhabdomyosarcoma, and three for bladder cancer. Four leukemia patients received additional radiation therapy and bone marrow transplantation. The median interval from chemotherapy to TESE was 8.5 years. Testicular sperm was obtained in 30 patients (48%). Chemical pregnancy was achieved in 22 patients, and 17 live deliveries were achieved. We could not find any predictive factor of outcome of TESE-ICSI among patient backgrounds and pretreatment variables. CONCLUSION: Testicular sperm could be retrieved in 48% of persistent azoospermic men after chemotherapy, and could be used to achieve delivery of healthy children. Supported by: Testicular sperm could be retrieved in 48% of persistent azoospermic men after chemotherapy, and could be used to achieve delivery of healthy children.
also with poor embryo development, chromosomal abnormalities, and noncompetent pre-implantatory embryos. This is the first clinical prospective and randomized study, which evaluate the use of MACS-Anexin V to select the sperm sample without early apoptotic spermatozoa. After applying this technique in male subjects with severe teratozoospermia (<4%), we observed a benefit in the fertilization rate and embryo development either for IVF or ICSI compared with swimp up. However, a bigger sample size is necessary to demonstrate a significant difference in the fertilization process.
P-972 Thursday, October 17, 2013 COMPARISON OF DNA FRAGMENTATION LEVELS IN SPERMIOGENESIS OF AZOOSPERMIC PATIENTS (OBSTRUCTIVE VS. NO OBSTRUCTIVE). C. Alvarez Sedo, F. Fulco, M. Lavolpe, H. Uriondo, F. Aguirre, G. Alvarez. CEGYR - Reproductive Medicine, Capital Federal, Buenos Aires, Argentina. OBJECTIVE: To compare DNA fragmentation levels in spermiogenesis from biopsies of obstructive vs. non obstructive azoospermic patients. DESIGN: Prospective and comparative study. MATERIALS AND METHODS: We included testicular biopsies of 35 azoospermic patients (no spermatozoa after at two spermograms with centrifugation). Testicular surgeries were performed between January 2011 - October 2012. Samples were divided in two groups: A) Obstructive azoospermia (N¼18) (16 for agenesis of the vas deferens and 2 for vasectomy) and B) Non-obstructive azoospermia (N¼17). Enzymatic digestion of tissue (hyaluronidase and trypsin) was performed to obtain spermatogenic isolated cells. These were fixed for subsequent immunocytochemistry. Acrosin protein was immunodetected and DNA staining (Hoechst), served as markers of the different stages of spermiogenesis: round spermatids, elongated and spermatozoa. In the same sample, TUNEL technique was applied for assessing DNA fragmentation. The statistical analysis employed was a student T-test for independent samples. RESULTS: The mean age among patients were similar, in group A was 35.43.2 (95%CI 33.7-37.4) and in group B: 38.76.4 (95%CI 32.844.7). The levels of DNA fragmentation in different stages are shown in the following table 1.
P-971 Thursday, October 17, 2013 HUMAN FERTILIZATION AND PRE-IMPLANTATORY EMBRYO QUALITY ENHANCEMENT APPLYING ANEXIN V MAGNETIC BINDING COLUMNS (MACS) ON IVF/ICSI INFERTILE COUPLES. G. Barroso,a,d C. Valdespin,a R. Avila,a A. Colin,a,d G. Estrada,c S. Oehninger.b aHealth Research, Nascere Reproductive Center, Mexico, Mexico; bThe Jones Institute for Reproductive Medicine, Eastern Virginia Medical School, Norfolk, VA; cResearch Division, Instituto Nacional de Perinatologia, Mexico, Mexico; dReproductive Health Research, The American British Cowdray Medical Center, Mexico, Mexico. OBJECTIVE: To compare the fertilization rate and embryo development on IVF/ICSI techniques when standard swimp up and MACS are applied on semen samples according to sperm morphology. DESIGN: Clinical controlled prospective trial. MATERIALS AND METHODS: A total of 387 oocytes on metaphase II were assigned into four groups according to the sperm preparation method and the insemination technique, as follows: group A (swimp up + ICSI), group B (MACS+ ICSI), group C (swimp up+ IVF), group D (MACS + IVF), on each group sperm morphology was evaluated according to Kruger strict criteria: 1-4%, 5-10%,R11% of normal sperm forms. Fertilization rate and embryo development, including: i) embryo division (number of cells), ii) fragmentation percentage and iii) multinucleation in day 2 and 3 were analyzed. Kruskal-Wallis and X2 test were performed for statistical analyses. RESULTS: Patients demographics were similar, significant differences were observed on the fertilization rate between group C vs A P¼0.0009, no significant differences between the other groups A vs B (84% vs 87%; P¼0.06), C vs D (63% vs 70% P¼0.23) were found. The fertilization rate in patients with sperm morphology <4% (strict Kruger criteria) was: 78%, 85%, 36% and 53% for group A, B, C and D respectively, which was statistically significant P>0.05. CONCLUSION: Male infertility derived from early and late apoptotic process has been associated with not only an anomalous fertilization process but
S428
ASRM Abstracts
Stages
Obstructive azoospermia (A)
Non obstructive azoospermia (B)
XSD (IC95%)
XSD (IC95%)
p
Round 31.0 6.4 (24.3-37.7) 53.3 14.3 (42.6-64.7) 0.08 Spermatids Elongated 15.0 4.9 (9.7-20.2) 30.9 12.7 (21.1-40.7) 0.03 Spermatid Spermatozoa 18.6 3.8 (14.2-22.1) 40.1 18.4 (26.6-54.9) 0.003
CONCLUSION: These results suggest that the failure in spermatogenesis process (Group B), may be related to greater impairment of DNA integrity. These data could be relevant for ICSI outcomes where sperm biopsies are used. Supported by: CEGYR Foundation.
P-973 Thursday, October 17, 2013 REPETATIVE PERCUTANEOUS EPIDIDYMAL SPERM ASPIRATIONS (PESA’S) IN A RAT MODEL RESULTED IN IMMEDIATE ASTHENOSPERMIA AND SIGNIFICANT INFLAMMATORY CHANGES. Y. Zhang,a K. R. Chohan,b S. K. Landas,b J. Reeder,a a a J. C. Trussell. Urology, Upstate University Hospital, Syracuse, NY; bPathology, Upstate University Hospital, Syracuse, NY. OBJECTIVE: In azoospermia, choosing a sperm retrieval method for intracytoplasmic sperm injection (ICSI) depends primarily on the preference and expertise of both the urologist and reproductive endocrinologist. There is insufficient evidence to suggest which method of sperm harvesting (PESA verses testis biopsy/aspiration) optimizes ICSI outcomes. Generally, PESA is attempted first. Not uncommonly, multiple PESA’s are necessary
Vol. 100, No. 3, Supplement, September 2013
also with poor embryo development, chromosomal abnormalities, and noncompetent pre-implantatory embryos. This is the first clinical prospective and randomized study, which evaluate the use of MACS-Anexin V to select the sperm sample without early apoptotic spermatozoa. After applying this technique in male subjects with severe teratozoospermia (<4%), we observed a benefit in the fertilization rate and embryo development either for IVF or ICSI compared with swimp up. However, a bigger sample size is necessary to demonstrate a significant difference in the fertilization process.
P-972 Thursday, October 17, 2013 COMPARISON OF DNA FRAGMENTATION LEVELS IN SPERMIOGENESIS OF AZOOSPERMIC PATIENTS (OBSTRUCTIVE VS. NO OBSTRUCTIVE). C. Alvarez Sedo, F. Fulco, M. Lavolpe, H. Uriondo, F. Aguirre, G. Alvarez. CEGYR - Reproductive Medicine, Capital Federal, Buenos Aires, Argentina. OBJECTIVE: To compare DNA fragmentation levels in spermiogenesis from biopsies of obstructive vs. non obstructive azoospermic patients. DESIGN: Prospective and comparative study. MATERIALS AND METHODS: We included testicular biopsies of 35 azoospermic patients (no spermatozoa after at two spermograms with centrifugation). Testicular surgeries were performed between January 2011 - October 2012. Samples were divided in two groups: A) Obstructive azoospermia (N¼18) (16 for agenesis of the vas deferens and 2 for vasectomy) and B) Non-obstructive azoospermia (N¼17). Enzymatic digestion of tissue (hyaluronidase and trypsin) was performed to obtain spermatogenic isolated cells. These were fixed for subsequent immunocytochemistry. Acrosin protein was immunodetected and DNA staining (Hoechst), served as markers of the different stages of spermiogenesis: round spermatids, elongated and spermatozoa. In the same sample, TUNEL technique was applied for assessing DNA fragmentation. The statistical analysis employed was a student T-test for independent samples. RESULTS: The mean age among patients were similar, in group A was 35.43.2 (95%CI 33.7-37.4) and in group B: 38.76.4 (95%CI 32.844.7). The levels of DNA fragmentation in different stages are shown in the following table 1.
P-971 Thursday, October 17, 2013 HUMAN FERTILIZATION AND PRE-IMPLANTATORY EMBRYO QUALITY ENHANCEMENT APPLYING ANEXIN V MAGNETIC BINDING COLUMNS (MACS) ON IVF/ICSI INFERTILE COUPLES. G. Barroso,a,d C. Valdespin,a R. Avila,a A. Colin,a,d G. Estrada,c S. Oehninger.b aHealth Research, Nascere Reproductive Center, Mexico, Mexico; bThe Jones Institute for Reproductive Medicine, Eastern Virginia Medical School, Norfolk, VA; cResearch Division, Instituto Nacional de Perinatologia, Mexico, Mexico; dReproductive Health Research, The American British Cowdray Medical Center, Mexico, Mexico. OBJECTIVE: To compare the fertilization rate and embryo development on IVF/ICSI techniques when standard swimp up and MACS are applied on semen samples according to sperm morphology. DESIGN: Clinical controlled prospective trial. MATERIALS AND METHODS: A total of 387 oocytes on metaphase II were assigned into four groups according to the sperm preparation method and the insemination technique, as follows: group A (swimp up + ICSI), group B (MACS+ ICSI), group C (swimp up+ IVF), group D (MACS + IVF), on each group sperm morphology was evaluated according to Kruger strict criteria: 1-4%, 5-10%,R11% of normal sperm forms. Fertilization rate and embryo development, including: i) embryo division (number of cells), ii) fragmentation percentage and iii) multinucleation in day 2 and 3 were analyzed. Kruskal-Wallis and X2 test were performed for statistical analyses. RESULTS: Patients demographics were similar, significant differences were observed on the fertilization rate between group C vs A P¼0.0009, no significant differences between the other groups A vs B (84% vs 87%; P¼0.06), C vs D (63% vs 70% P¼0.23) were found. The fertilization rate in patients with sperm morphology <4% (strict Kruger criteria) was: 78%, 85%, 36% and 53% for group A, B, C and D respectively, which was statistically significant P>0.05. CONCLUSION: Male infertility derived from early and late apoptotic process has been associated with not only an anomalous fertilization process but
S428
ASRM Abstracts
Stages
Obstructive azoospermia (A)
Non obstructive azoospermia (B)
XSD (IC95%)
XSD (IC95%)
p
Round 31.0 6.4 (24.3-37.7) 53.3 14.3 (42.6-64.7) 0.08 Spermatids Elongated 15.0 4.9 (9.7-20.2) 30.9 12.7 (21.1-40.7) 0.03 Spermatid Spermatozoa 18.6 3.8 (14.2-22.1) 40.1 18.4 (26.6-54.9) 0.003
CONCLUSION: These results suggest that the failure in spermatogenesis process (Group B), may be related to greater impairment of DNA integrity. These data could be relevant for ICSI outcomes where sperm biopsies are used. Supported by: CEGYR Foundation.
P-973 Thursday, October 17, 2013 REPETATIVE PERCUTANEOUS EPIDIDYMAL SPERM ASPIRATIONS (PESA’S) IN A RAT MODEL RESULTED IN IMMEDIATE ASTHENOSPERMIA AND SIGNIFICANT INFLAMMATORY CHANGES. Y. Zhang,a K. R. Chohan,b S. K. Landas,b J. Reeder,a a a J. C. Trussell. Urology, Upstate University Hospital, Syracuse, NY; bPathology, Upstate University Hospital, Syracuse, NY. OBJECTIVE: In azoospermia, choosing a sperm retrieval method for intracytoplasmic sperm injection (ICSI) depends primarily on the preference and expertise of both the urologist and reproductive endocrinologist. There is insufficient evidence to suggest which method of sperm harvesting (PESA verses testis biopsy/aspiration) optimizes ICSI outcomes. Generally, PESA is attempted first. Not uncommonly, multiple PESA’s are necessary
Vol. 100, No. 3, Supplement, September 2013