title: Separation of cryopreserved human semen using Sephadex columns, washing, or Percoll gradients.
abstract: The following methods were evaluated for their ability to separate motile cryopreserved sperm from semen after thawing: single washing, Percoll separation followed by a single washing, and Sephadex column separation. For Sephadex separation, washing, and Percoll separation, percent recovery of motile sperm was 65%, 76%, and 28%, and motility was 81%, 39%, and 60%, respectively. Percoll separation and washing were the best methods for removing seminal constituents, but sperm velocity and linearity were lower after Percoll separation and washing than after Sephadex separation. During 3 hours of incubation, there was an additional decrease in the motility, viability (exclusion of supravital dye), velocity, linearity, and intact acrosomes of Percoll-separated sperm, indicating that Percoll separation may not be suitable for cryopreserved sperm. Motile, washed sperm also had lower velocities and higher spontaneous acrosome reactions than Sephadex-separated sperm, but velocity and linearity were maintained during incubation. When semen was separated with Sephadex followed by washing, motility was well maintained (84%). The Sephadex method is a promising technique for selecting and concentrating motile cryopreserved sperm.
authors.affiliation: Department of Obstetrics and Gynecology, School of Medicine, University of California, Davis 95616.
authors.forename: E Z
authors.initials: EZ
authors.lastname: Drobnis
authors.forename: C Q
authors.initials: CQ
authors.lastname: Zhong
authors.forename: J W
authors.initials: JW
authors.lastname: Overstreet
mesh_terms: D002469:Cell Separation / Q000379:methods*; D002470:Cell Survival; D002499:Centrifugation, Density Gradient; D002853:Chromatography, Liquid; D015925:Cryopreservation; D005069:Evaluation Studies as Topic; D006801:Humans; D008297:Male; D011205:Povidone; D012661:Semen; D012662:Semen Preservation; D012822:Silicon Dioxide; D013081:Sperm Motility; D013094:Spermatozoa
chemical_list: C016039:Percoll; D012822:Silicon Dioxide; D011205:Povidone