Aim: To compare the recovery rate of morphologically normal and chromatin condensed spermatozoa from native se-men samples using the SpermPrep^(TM) filtration columns and Percoll gradient centrifugation and to determi...Aim: To compare the recovery rate of morphologically normal and chromatin condensed spermatozoa from native se-men samples using the SpermPrep^(TM) filtration columns and Percoll gradient centrifugation and to determine the influenceof the two processing techniques on fertilization and pregnancy rates in an IVF-ET program. Methods: Sixteen se-men samples obtained from patient's husband were included in this study. Each was divided into two aliquots. The firstaliquot was processed with SpermPrep^(TM) filtration columns and the second, Percoll gradient centrifugation. Smears weremade before and after semen processing with both methods for the evaluation of chromatin condensation (chromomycineCMA3) as well as morphology (strict criteria) of spermatozoa. One hundred and seventy oocytes were retrieved fromthe patients and the oocytes from each patient were subdivided into two sets : one set was inseminated using spermatozoaprocessed with SpermPrep^(TM) and the other inseminated after semen processing with Percoll gradient centrifugation. Re-sults: The Percoll method yielded a significantly higher percentage of chromatin condensed (90.8 ±6.5% vs 82.3±8.8%, P = 0.017) and morphologically normal spermatozoa (12.9±7.4% vs 6.9±4.8%, P =0.001) in com-parison to SpermPrep^(TM). Whereas, sperm count recovery rate was significantly higher after the use of SpermPrep^(TM) thanafter the Percoll gradient centrifugation. The fertilization rate was similar between the two methods. Conclusion:Semen processing with Percoll should be recommended for intracytoplasmic sperm injection as the natural selection isbypassed and the SpermPrep^(TM) technique could be recommended for IVF and IUI programs as the sperm concentrationplays a more significant role in these procedures.展开更多
This was a prospective multicenter study aiming at comparing the efficiency of sperm selection by density gradient centrifugation (DGC) in reducing sperm DNA fragmentation (SDF) in different ART centers. The study was...This was a prospective multicenter study aiming at comparing the efficiency of sperm selection by density gradient centrifugation (DGC) in reducing sperm DNA fragmentation (SDF) in different ART centers. The study was designed using 290 semen samples collected from 10 different ART centers performing artificial insemination, in vitro fertilization and blind assessment of SDF at the University facilities. The results showed that while there was a significant reduction in the SDF levels in sperm isolated from the gradient centrifuged pellet (DGC) compared to neat semen samples (NSS), there was also significant inter-center variability in the efficiency to reduce SDF values by DGC (78.5% to 29.2%). Surprisingly, for some patients, the level of SDF actually increased following sperm selection. The main conclusions derived from this study were that 1) isolation of sperm from the gradient pellet by DGC must be performed using validated, optimized protocols;2) routine comparison of SDF values in NSS semen and in processed sperm after DGC or swim-up must be recommended as part of the internal quality control (QC) of ART laboratories to test the efficacy of sperm processing;and 3) SDF values in processed spermatozoa should be obtained to compare with the pregnancy rate when insemination or fertilization is about to be performed, otherwise, attempts to predict pregnancy outcome from SDF could be biased or are essentially meaningless.展开更多
文摘Aim: To compare the recovery rate of morphologically normal and chromatin condensed spermatozoa from native se-men samples using the SpermPrep^(TM) filtration columns and Percoll gradient centrifugation and to determine the influenceof the two processing techniques on fertilization and pregnancy rates in an IVF-ET program. Methods: Sixteen se-men samples obtained from patient's husband were included in this study. Each was divided into two aliquots. The firstaliquot was processed with SpermPrep^(TM) filtration columns and the second, Percoll gradient centrifugation. Smears weremade before and after semen processing with both methods for the evaluation of chromatin condensation (chromomycineCMA3) as well as morphology (strict criteria) of spermatozoa. One hundred and seventy oocytes were retrieved fromthe patients and the oocytes from each patient were subdivided into two sets : one set was inseminated using spermatozoaprocessed with SpermPrep^(TM) and the other inseminated after semen processing with Percoll gradient centrifugation. Re-sults: The Percoll method yielded a significantly higher percentage of chromatin condensed (90.8 ±6.5% vs 82.3±8.8%, P = 0.017) and morphologically normal spermatozoa (12.9±7.4% vs 6.9±4.8%, P =0.001) in com-parison to SpermPrep^(TM). Whereas, sperm count recovery rate was significantly higher after the use of SpermPrep^(TM) thanafter the Percoll gradient centrifugation. The fertilization rate was similar between the two methods. Conclusion:Semen processing with Percoll should be recommended for intracytoplasmic sperm injection as the natural selection isbypassed and the SpermPrep^(TM) technique could be recommended for IVF and IUI programs as the sperm concentrationplays a more significant role in these procedures.
基金supported with public funding(Spanish Ministry of Science and Technology(MCYT:BFU2010-16738).
文摘This was a prospective multicenter study aiming at comparing the efficiency of sperm selection by density gradient centrifugation (DGC) in reducing sperm DNA fragmentation (SDF) in different ART centers. The study was designed using 290 semen samples collected from 10 different ART centers performing artificial insemination, in vitro fertilization and blind assessment of SDF at the University facilities. The results showed that while there was a significant reduction in the SDF levels in sperm isolated from the gradient centrifuged pellet (DGC) compared to neat semen samples (NSS), there was also significant inter-center variability in the efficiency to reduce SDF values by DGC (78.5% to 29.2%). Surprisingly, for some patients, the level of SDF actually increased following sperm selection. The main conclusions derived from this study were that 1) isolation of sperm from the gradient pellet by DGC must be performed using validated, optimized protocols;2) routine comparison of SDF values in NSS semen and in processed sperm after DGC or swim-up must be recommended as part of the internal quality control (QC) of ART laboratories to test the efficacy of sperm processing;and 3) SDF values in processed spermatozoa should be obtained to compare with the pregnancy rate when insemination or fertilization is about to be performed, otherwise, attempts to predict pregnancy outcome from SDF could be biased or are essentially meaningless.
