title: Combination of swim-up and density gradient separation methods effectively eliminate DNA damaged sperm.
abstract: OBJECTIVE
  The aim of this experimental prospective study was to investigate the efficacy of single and combination sperm wash methods for their ability to isolate DNA intact spermatozoa.
  
  
  METHODS
  Sperm DNA damage was introduced by local testicular irradiation in male mice and the extent of damage was quantified by comet assay. The spermatozoa were subjected to single (swim up or density gradient method) and also a combination of sperm wash techniques. The DNA integrity in various sub-fractions of wash techniques was evaluated.
  
  
  RESULTS
  The amount of DNA damaged sperm did not differ between individual fractions when single wash technique was applied. However, a combination of density gradient and swim-up techniques significantly reduced (p<0.01) the number of DNA damaged sperm in the final population.
  
  
  CONCLUSIONS
  The combination of density gradient separation and swim-up method is effective in eliminating DNA damaged spermatozoa.
authors.affiliation: Clinical Embryology, Division of Reproductive Medicine, Kasturba Medical College, Manipal University, Manipal, India.
authors.forename: Surveen
authors.initials: S
authors.lastname: Ghumman
authors.affiliation: Clinical Embryology, Division of Reproductive Medicine, Kasturba Medical College, Manipal University, Manipal, India.
authors.forename: Satish Kumar
authors.initials: SK
authors.lastname: Adiga
authors.affiliation: Clinical Embryology, Division of Reproductive Medicine, Kasturba Medical College, Manipal University, Manipal, India.
authors.forename: Dinesh
authors.initials: D
authors.lastname: Upadhya
authors.affiliation: Clinical Embryology, Division of Reproductive Medicine, Kasturba Medical College, Manipal University, Manipal, India.
authors.forename: Guruprasad
authors.initials: G
authors.lastname: Kalthur
authors.affiliation: Clinical Embryology, Division of Reproductive Medicine, Kasturba Medical College, Manipal University, Manipal, India.
authors.forename: Varshini
authors.initials: V
authors.lastname: Jayaraman
authors.affiliation: Department of Radiobiology and Toxicology, Manipal Life Science Centre, Manipal, India.
authors.forename: Satish Bola
authors.initials: SB
authors.lastname: Rao
authors.affiliation: Clinical Embryology, Division of Reproductive Medicine, Kasturba Medical College, Manipal University, Manipal, India.
authors.forename: Pratap
authors.initials: P
authors.lastname: Kumar
keywords: DNA damage; Sperm wash; comet assay; density gradient separation; swim-up