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.
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.
Authors: U Punjabi; H Van Mulders; I Goovaerts; K Peeters; E Roelant; D De Neubourg Journal: J Assist Reprod Genet Date: 2019-05-14 Impact factor: 3.412
Authors: Reza Nosrati; Percival J Graham; Biao Zhang; Jason Riordon; Alexander Lagunov; Thomas G Hannam; Carlos Escobedo; Keith Jarvi; David Sinton Journal: Nat Rev Urol Date: 2017-10-31 Impact factor: 14.432
Authors: Karl R Hansen; Jennifer D Peck; R Matthew Coward; Robert A Wild; J C Trussell; Stephen A Krawetz; Michael P Diamond; Richard S Legro; Christos Coutifaris; Ruben Alvero; Randal D Robinson; Peter Casson; Gregory M Christman; Nanette Santoro; Heping Zhang Journal: Hum Reprod Date: 2020-06-01 Impact factor: 6.353