Fulltext

The use of two density gradient centrifugation techniques and the swim-up method to separate spermatozoa with chromatin and nuclear DNA anomalies

Sakkas, D. ; Manicardi, G.C. ; Tomlinson, M. ; Mandrioli, M. ; Bizzaro, D. ; Bianchi, P.G. ; Bianchi, U.

In: Human Reproduction, 2000, vol. 15, no. 5, p. 1112-1116

Ajouter à la liste personnelle
    Titre
    • The use of two density gradient centrifugation techniques and the swim-up method to separate spermatozoa with chromatin and nuclear DNA anomalies
    Auteur
    • Sakkas, D.. Assisted Conception Unit, Birmingham Women's Hospital, Edgbaston, Birmingham B15 2TG, UK
    • Manicardi, G.C.. Department of Animal Biology, University of Modena, Italy and
    • Tomlinson, M.. Assisted Conception Unit, Birmingham Women's Hospital, Edgbaston, Birmingham B15 2TG, UK
    • Mandrioli, M.. Department of Animal Biology, University of Modena, Italy and
    • Bizzaro, D.. Department of Animal Biology, University of Modena, Italy and
    • Bianchi, P.G.. Clinic of Sterility, Department of Obstetrics and Gynaecology, University Hospital of Geneva, Switzerland
    • Bianchi, U.. Department of Animal Biology, University of Modena, Italy and
    Type de document
    • Postprint
    Langue
    • Anglais
    Publié dans
    • Human Reproduction, 2000, vol. 15, no. 5, p. 1112-1116. Oxford University Press
    Autre version électronique
    Identifiant OAI-PMH
    • oai:doc.rero.ch:289227
    Summary
    Human semen is heterogeneous in quality, not only between males but also within a single ejaculate. Differences in quality are evident, both when examining the classical parameters of sperm number, motility and morphology and in the integrity of the sperm nucleus. The aim of this study was to determine the efficiency of the PureSperm®, Percoll® and swim-up preparation techniques to eliminate spermatozoa with nuclear anomalies. Semen samples were collected, washed and one part of the semen spread on a slide, the remainder was prepared using the swim-up, PureSperm® or Percoll® techniques. Spermatozoa from different fractions were fixed on slides and assessed. Sperm samples (n) from different men were stained using the chromomycin A3 (CMA3) fluorochrome, which indirectly demonstrates a decreased presence of protamine (n = 31 for swim-up; n = 45 for PureSperm®; n = 39 for Percoll®). Spermatozoa prepared using PureSperm® (n = 35) and Percoll® (n = 37) were also examined for the presence of endogenous DNA nicks. Good quality spermatozoa should not possess DNA nicks and not stain (i.e. fluoresce) with CMA3. When prepared using the swim-up technique the spermatozoa recovered showed no significant improvement with the CMA3 staining. When spermatozoa were prepared using the PureSperm® and Percoll® techniques, a significant (P < 0.001) decrease in both CMA3 positivity and DNA strand breakage was observed. These results indicate that both the PureSperm® and Percoll® techniques can enrich the sperm population by separating out those with nicked DNA and with poorly condensed chromatin