A Zini1, V Mak, D Phang, K Jarvi. 1. Department of Surgery, Mount Sinai Hospital, University of Toronto, Ontario, Canada. azini@mtsinai.on.ca
Abstract
OBJECTIVE: To examine the effect of standard Percoll density-gradient centrifugation on human sperm DNA denaturation. DESIGN: Prospective, observational study. SETTING: University-based infertility clinic. PATIENT(S): Twenty-five nonazoospermic men. INTERVENTION(S): Semen samples (n = 25) were obtained from consecutively seen nonazoospermic men presenting for infertility evaluation. Samples were processed by two-layer and four-layer Percoll density gradients. Sperm motility and sperm chromatin structure (evaluated by flow cytometry analysis of acridine orange-treated spermatozoa) were monitored before and after semen processing. Sperm chromatin integrity was expressed as the percentage of spermatozoa that demonstrated denatured DNA. MAIN OUTCOME MEASURE(S): Sperm motility and DNA integrity. RESULT(S): Mean sperm motility improved significantly after processing with two-layer and four-layer Percoll gradients compared with whole semen (54% and 57% motility versus 44% motility, respectively). In contrast, the percentage of sperm with denatured DNA increased after processing with two-layer and four-layer Percoll gradients compared with whole semen (34% and 32% versus 18%, respectively). CONCLUSION(S): Our data demonstrate that the improvement seen in sperm motility after Percoll processing is not associated with a similar improvement in sperm DNA integrity. These data suggest that we reexamine current sperm processing techniques to minimize sperm DNA damage and the potential transmission of genetic mutations in assisted reproductive cycles.
OBJECTIVE: To examine the effect of standard Percoll density-gradient centrifugation on human sperm DNA denaturation. DESIGN: Prospective, observational study. SETTING: University-based infertility clinic. PATIENT(S): Twenty-five nonazoospermic men. INTERVENTION(S): Semen samples (n = 25) were obtained from consecutively seen nonazoospermic men presenting for infertility evaluation. Samples were processed by two-layer and four-layer Percoll density gradients. Sperm motility and sperm chromatin structure (evaluated by flow cytometry analysis of acridine orange-treated spermatozoa) were monitored before and after semen processing. Sperm chromatin integrity was expressed as the percentage of spermatozoa that demonstrated denatured DNA. MAIN OUTCOME MEASURE(S): Sperm motility and DNA integrity. RESULT(S): Mean sperm motility improved significantly after processing with two-layer and four-layer Percoll gradients compared with whole semen (54% and 57% motility versus 44% motility, respectively). In contrast, the percentage of sperm with denatured DNA increased after processing with two-layer and four-layer Percoll gradients compared with whole semen (34% and 32% versus 18%, respectively). CONCLUSION(S): Our data demonstrate that the improvement seen in sperm motility after Percoll processing is not associated with a similar improvement in sperm DNA integrity. These data suggest that we reexamine current sperm processing techniques to minimize sperm DNA damage and the potential transmission of genetic mutations in assisted reproductive cycles.
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