title: Evaluation of conventional semen parameters, intracellular reactive oxygen species, DNA fragmentation and dysfunction of mitochondrial membrane potential after semen preparation techniques: a flow cytometric study.
abstract: OBJECTIVE
  The aim of this study was to evaluate the conventional sperm parameters, level of intracellular reactive oxygen species (ROS), DNA fragmentation (DF) and dysfunction of mitochondrial membrane potential (MMP) after semen preparation techniques with flow cytometry.
  
  
  METHODS
  Semen samples were obtained from 28 men with normal semen analysis according to WHO (world health organization). Each was divided into three equal parts for processing with routine techniques: conventional swim up (CSW), direct swim up (DSW) and density gradient centrifugation (DGC). The conventional sperm parameters were evaluated with computer-assisted sperm analyzer (CASA) and the level of intracellular ROS, dysfunction of MMP and DF were determined with flow cytometry procedure.
  
  
  RESULTS
  Conventional sperm parameters such as motility, progressive motility and normal morphology increased after sperm processing by CSW and DGC compared to DSW. A significant increase in intracellular H₂O₂ (p < 0.05) was demonstrated in the CSW versus DSW technique, while processed sperm by the DSW procedure showed a significant increase in the percentage of dysfunction of MMP and intracellular O₂(•-) (p < 0.05) when compared with CSW and DGC techniques. Additionally, a high mean of DF (p < 0.05) was observed in the DGC technique as compared to CSW.
  
  
  CONCLUSIONS
  Data from flow cytometry study demonstrated that intracellular H₂O₂ and DF increased after CSW and DGC processing techniques, respectively, whereas the level of intracellular O₂(•-) and dysfunction of MMP only increased after the DSW processing technique.
authors.affiliation: Department of Embryology at Reproductive Biomedicine Research Center, Royan Institute for Reproductive Biomedicine, ACECR, P. O. 19395-4644, Tehran, Iran, lrashki@yahoo.com.
authors.forename: Leila Rashki
authors.initials: LR
authors.lastname: Ghaleno
authors.forename: Mojtaba Rezazadeh
authors.initials: MR
authors.lastname: Valojerdi
authors.forename: Ehsan
authors.initials: E
authors.lastname: Janzamin
authors.forename: Mohammad
authors.initials: M
authors.lastname: Chehrazi
authors.forename: Mina
authors.initials: M
authors.lastname: Sharbatoghli
authors.forename: Reza Salman
authors.initials: RS
authors.lastname: Yazdi
mesh_terms: D053938:DNA Fragmentation; D005434:Flow Cytometry; D006801:Humans; D006861:Hydrogen Peroxide / Q000378:metabolism; D008297:Male; D053078:Membrane Potential, Mitochondrial / Q000502:physiology*; D017382:Reactive Oxygen Species / Q000378:metabolism*; D012661:Semen / Q000166:cytology* / Q000502:physiology; D055101:Semen Analysis; D013094:Spermatozoa / Q000166:cytology / Q000502:physiology
chemical_list: D017382:Reactive Oxygen Species; D006861:Hydrogen Peroxide