A wide variety of techniques for the preparation of sperm are currently available, of which the most commonly employed are densitygradient centrifugation （DGC） and swim-up （SUP）. To date, these methods appear to b...A wide variety of techniques for the preparation of sperm are currently available, of which the most commonly employed are densitygradient centrifugation （DGC） and swim-up （SUP）. To date, these methods appear to be effective in selecting functional sperm for assisted reproduction techniques （ART）, but they may have negative effects on sperm DNA. In this study, the ability of these semen processing techniques to eliminate spermatozoa containing single- and double-strand DNA damage was assessed by the two-tailed comet assay and the sperm chromatin dispersion test in 157 semen samples from patients seeking assisted reproduction treatment. Our results indicated that SUP and DGC are equally efficient in eliminating spermatozoa containing double-strand DNA damage and sperm with highly damaged （degraded） DNA, as characterized by the presence of both single- and double-strand DNA breaks. However, DGC is more efficient than SUP in selecting spermatozoa that are free from single-strand DNA damage. Future studies should characterise the importance of the various types of DNA damage and examine the sperm processing protocols used in each laboratory to determine their ability to eliminate DNA damage and hence, prevent the potential transmission of genetic mutations via ART.展开更多
Assisted reproductive technologies invoIving the use of spermatozoa and eggs for in vitro fertilization(IVF)have come as the solution for many infertile couples to become parents.However,in some cases,the use of ejacu...Assisted reproductive technologies invoIving the use of spermatozoa and eggs for in vitro fertilization(IVF)have come as the solution for many infertile couples to become parents.However,in some cases,the use of ejaculated spermatozoa delivers poor IVF performance.Some studies have suggested the use of testicular spermatozoa in severe male in fertility cases,but no guideli nes regarding their utilization are currently available.In the present study,we found the mRNA protamine 1/protamine 2(P1/P2)ratio to be a valuable biomarker of poor sperm function that could be used as a diagnostic key for the identification of cases that would benefit from the use of testicular spermatozoa.A total of 23 couples undergoing egg donation cycles with at least one previous cycle failure were studied.All couples underwent two consecutive intracytoplasmic sperm injection(ICSI)cycles with either ejaculated or testicular spermatozoa(TESA).The sperm mRNA P1/P2 ratio,fertilization rate,blastocyst rate,and pregnancy and live birth rate were compared.Results showed improved ICSI and clinical outcomes in cycles with testicular spermatozoa in men with altered mRNA P1/P2 ratios.TESA cycles presented significantly higher rates of fertilization(mean±standard deviation:76.1%±15.1%vs 65.5%±18.8%),blastocyst formation(55.0%±20.3%vs 30.8%±23.8%),and good morphological quality blastocyst(28.9%±22.9%vs 13.5%±17.9%)and also improvements on pregnancy(60.9%vs 0%)and healthy birth rates(56.5%vs 0%)than EJACULATE cycles.The results described here suggest that in patients with previous IVF/ICSI failures and aberrant mRNA protamine ratios,the use of testicular spermatozoa may be a good alternative to improve clinical outcomes.展开更多
文摘A wide variety of techniques for the preparation of sperm are currently available, of which the most commonly employed are densitygradient centrifugation （DGC） and swim-up （SUP）. To date, these methods appear to be effective in selecting functional sperm for assisted reproduction techniques （ART）, but they may have negative effects on sperm DNA. In this study, the ability of these semen processing techniques to eliminate spermatozoa containing single- and double-strand DNA damage was assessed by the two-tailed comet assay and the sperm chromatin dispersion test in 157 semen samples from patients seeking assisted reproduction treatment. Our results indicated that SUP and DGC are equally efficient in eliminating spermatozoa containing double-strand DNA damage and sperm with highly damaged （degraded） DNA, as characterized by the presence of both single- and double-strand DNA breaks. However, DGC is more efficient than SUP in selecting spermatozoa that are free from single-strand DNA damage. Future studies should characterise the importance of the various types of DNA damage and examine the sperm processing protocols used in each laboratory to determine their ability to eliminate DNA damage and hence, prevent the potential transmission of genetic mutations via ART.
基金The authors thank all the patients for consenting to participate in this study.Furthermore,the technical assistance of Barbara Frohlich and Mareike Buch-Heberling is gratefully acknowledged.KS was supported by a Research Grant from the University Medical Center Giessen and Marburg(UKGM,project 29/2015GI).This research did not receive any other specific grant from funding agencies in the public,commercial,or not-for-profit sectors.
文摘Assisted reproductive technologies invoIving the use of spermatozoa and eggs for in vitro fertilization(IVF)have come as the solution for many infertile couples to become parents.However,in some cases,the use of ejaculated spermatozoa delivers poor IVF performance.Some studies have suggested the use of testicular spermatozoa in severe male in fertility cases,but no guideli nes regarding their utilization are currently available.In the present study,we found the mRNA protamine 1/protamine 2(P1/P2)ratio to be a valuable biomarker of poor sperm function that could be used as a diagnostic key for the identification of cases that would benefit from the use of testicular spermatozoa.A total of 23 couples undergoing egg donation cycles with at least one previous cycle failure were studied.All couples underwent two consecutive intracytoplasmic sperm injection(ICSI)cycles with either ejaculated or testicular spermatozoa(TESA).The sperm mRNA P1/P2 ratio,fertilization rate,blastocyst rate,and pregnancy and live birth rate were compared.Results showed improved ICSI and clinical outcomes in cycles with testicular spermatozoa in men with altered mRNA P1/P2 ratios.TESA cycles presented significantly higher rates of fertilization(mean±standard deviation:76.1%±15.1%vs 65.5%±18.8%),blastocyst formation(55.0%±20.3%vs 30.8%±23.8%),and good morphological quality blastocyst(28.9%±22.9%vs 13.5%±17.9%)and also improvements on pregnancy(60.9%vs 0%)and healthy birth rates(56.5%vs 0%)than EJACULATE cycles.The results described here suggest that in patients with previous IVF/ICSI failures and aberrant mRNA protamine ratios,the use of testicular spermatozoa may be a good alternative to improve clinical outcomes.
