Prolonged Semen Incubation Alters the Biological Characteristics of Human Spermatozoa

• Published in: Iranian journal of medical sciences 2023-01, Vol.48 (S1), p.16
• Main Authors: Beigi, Sayed Abbas Datli, Khalili, Mohammad Ali, Nabi, Ali, Lorian, Keivan, Sabour, Mojdeh
• Format: Article
• Language: English
• Subjects: Chromatin; Spermatozoa
• ISSN: 0253-0716

Background: Semen incubation is a routine and crucial procedure in andrology laboratories. Therefore, paying attention to the incubation time in order to reduce possible damage to sperm parameters and other specific factors is noteworthy. The objective of this study was to assess the biological characteristics of human spermatozoa at different time intervals (0, 1, 1.5, and 2 hours) after incubation at 37 [degrees]C. Methods: 25 raw normozoospermic semen samples were incubated at 37 [degrees]C. Incubation was performed in four-time intervals of 0 (after liquefaction), 1, 1.5, and 2 hours. Samples were evaluated for sperm parameters at each time interval. Sperm motility was reported as a percentage of non-motile, non-progressive, and progressive motility. Sperm viability and morphology were assessed using Eosin-Nigrosin and Diff-Quik staining, respectively. Reactive oxygen species production, sperm mitochondria, chromatin, and acrosome reaction were also assessed. DNA fragmentation rate was evaluated according to halo formation around the sperm head. Results: The rate of progressive sperm motility decreased at 1.5 hours compared to 0 hour as well as 2 hours compared to the 1 hour and 0 hour. No significant changes were observed in sperm viability at any time intervals. Abnormal sperm morphology increased at 1.5 hours of incubation time. No significant changes were observed in DNA fragmentation at 1 hour compared to 0 hour, and 1.5 hours compared to 1 hour. However, a significant increase in DNA fragmentation was observed at 1.5 hours compared to 0 hour. Mitochondria membrane potential was decreased remarkably after 1 hour of incubation time. No significant differences were observed in acrosome reaction at 0, 1, 1.5, and 2 hours as well as malondialdehyde levels. Conclusion: The normozoospermic samples incubation before using in assisted reproductive technology process should be less than 1.5 hours to minimize the destructive effects of prolonged incubation time on general and specific sperm parameters. It is, therefore, recommended to perform sperm processing within the time frame of 1 hour of incubation. Keywords * Spermatozoa * DNA fragmentation * Acrosome reaction * Mitochondria * Membrane potentials