Effects of semen storage and separation techniques on sperm DNA fragmentation
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2010-12-01
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Motta, Eduardo L. A. 
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Objective: To determine the effect of semen storage and separation techniques on sperm DNA fragmentation.Design: Controlled clinical study.Setting: An assisted reproductive technology laboratory.Patient(s): Thirty normoozospermic semen samples obtained from patients undergoing infertility evaluation.Intervention(s): One aliquot from each sample was immediately prepared (control) for the sperm chromatin dispersion assay (SCD). Aliquots used to assess storage techniques were treated in the following ways: snap frozen by liquid nitrogen immersion, slow frozen with Tris-yolk buffer and glycerol, kept on ice for 24 hours or maintained at room temperature for 4 and 24 hours. Aliquots used to assess separation techniques were processed by the following methods: washed and centrifuged in media, swim-up from washed sperm pellet, density gradient separation, density gradient followed by swim-up. DNA integrity was then measured by SCD.Main Outcome Measure(s): DNA fragmentation as measured by SCD.Result(s): There was no significant difference in fragmentation among the snap frozen, slow frozen, and wet-ice groups. Compared to other storage methods short-term storage at room temperature did not impact DNA fragmentation yet 24 hours storage significantly increased fragmentation. Swim-up, density gradient and density gradient/swim-up had significantly reduced DNA fragmentation levels compared with washed semen. Postincubation, density gradient/swim-up showed the lowest fragmentation levels.Conclusion(s): the effect of sperm processing methods on DNA fragmentation should be considered when selecting storage or separation techniques for clinical use. (Fertil Steril (R) 2010;94:2626-30. (C) 2010 by American Society for Reproductive Medicine.)
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Fertility and Sterility. New York: Elsevier B.V., v. 94, n. 7, p. 2626-2630, 2010.