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Effects of temperature and storage time on the motility,viability, DNA integrity and apoptosis of processed human spermatozoa
Authors:Farhad Golshan Iranpour  Zohre Nateghian  Ralf Henkel  Gholam Reza Dashti
Institution:1. Department of Anatomical Sciences, Isfahan University of Medical Sciences, Isfahan, Iran

Saint Maryam Infertility Center, Shahid Beheshti Hospital, Isfahan University of Medical Sciences, Isfahan, Iran;2. Department of Anatomical Sciences, Isfahan University of Medical Sciences, Isfahan, Iran;3. Department of Medical Biosciences, University of the Western Cape, Bellville, South Africa

Abstract:The aim of this study was to evaluate motility, viability, DNA integrity and apoptosis of spermatozoa when washed semen samples were kept for up to 12 days at 4–6°C and 25°C. In this experimental study, 26 normozoospermic semen samples were washed twice in Modified Ham's F10 and resuspended in IVF fertilisation medium. Half of the specimens were stored at 4–6°C, and the other half was kept at 25°C for 12 days. The proportions of viable, motile, spermatozoa with double-stranded DNA and apoptotic spermatozoa were examined during storage time. Apoptosis was measured using annexin V-PI staining followed by flow cytometry. Results showed that sperm motility and viability decreased during 12 days of sample storage (p < .001). There was no significant difference between the two temperatures in terms of motility and viability for up to 2 days (p < .05). The percentage of spermatozoa with double-stranded DNA remained unchanged during the 12 days of storage at both temperatures (> .05). Although there was no difference between the two temperatures in terms of motility, viability and apoptosis during the first two days of storage, storage of spermatozoa at 4–6°C is better than storage for a longer period than storage at 25°C. Sperm DNA resisted against denaturation during storage.
Keywords:apoptosis  double-stranded DNA  semen preservation  sperm motility  spermatozoa
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