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Human Reproduction, Vol. 16, No. 10, 2160-2165, October 2001
© 2001 European Society of Human Reproduction and Embryology

Interrelationships between seminal parameters and sperm nuclear DNA damage before and after density gradient centrifugation: implications for assisted conception

M.J. Tomlinson1, O. Moffatt1, G.C. Manicardi2, D. Bizzaro2,3, M. Afnan1 and D. Sakkas4,5

1 Assisted Conception Unit, Birmingham Women's Hospital, UK, 2 Department of Animal Biology, University of Modena and Reggio Emilia, Italy, 3 Institute of Biology and Genetics, University of Ancona, Italy and 4 Department of Obstetrics and Gynaecology, Yale University School of Medicine, Connecticut, USA

BACKGROUND: With an increase in the use of assisted reproduction technologies the requirements of the diagnostic semen analysis are constantly changing. METHODS: Spermatozoa from patients undergoing IVF were analysed by examining the conventional semen parameters and DNA/chromatin integrity, using in-situ nick translation (NT) and the Chromomycin A3 fluorochrome, which indirectly demonstrates a decreased presence of protamine. Samples were examined before and after preparation using discontinuous density gradient centrifugation. RESULTS: Density gradient centrifugation enriched samples by improving the percentage of morphologically normal forms by 138% and sperm nuclear integrity by 450%. Sperm nuclear integrity as assessed by in-situ nick translation (NT) demonstrated a very clear relationship with sperm concentration, motility and morphology. Morphology correlated with fertilization rates of patients undergoing IVF, while NT values of the spermatozoa post-preparation were significantly lower in pregnant patients. CONCLUSIONS: We have demonstrated that along with the classical semen parameters, the assessment of nuclear integrity improves the characterization of the semen sample and may be used as a tool for allocating patients to specific assisted reproduction treatments.

Key words: semen parameters/sperm chromatin/sperm morphology/sperm nuclear DNA/sperm preparation

5 To whom correspondence should be addressed at: Department of Obstetrics and Gynecology, Yale University School of Medicine, 333 Cedar St., PO Box 208063, New Haven, CT 06520-8063, USA. E-mail: Denny.Sakkas{at}yale.edu


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