The effect of semen processing on sperm DNA integrity: comparison of two techniques using the novel Toluidine Blue Assay

Eur J Obstet Gynecol Reprod Biol. 2010 Aug;151(2):176-80. doi: 10.1016/j.ejogrb.2010.05.003. Epub 2010 Jun 8.

Abstract

Objective: The first goal of this study was to determine the effect that semen processing has on sperm DNA integrity. The second goal was to assess which processing technique (modified swim-up versus density gradient centrifugation) results in a superior sample. DNA integrity was measured using a novel Toluidine Blue Assay.

Study design: Side-by-side comparison.

Materials and methods: Raw semen samples were collected from thirty-two male individuals and scored for routine semen analysis. Prior to discarding the specimens identical aliquots were divided and processed by density gradient centrifugation and a modified swim-up technique. The Toluidine Blue Assay was used to analyze raw and processed samples.

Results: Both density gradient centrifugation and the modified swim-up improved DNA quality compared to the unprocessed sample. However, the modified swim-up technique proved superior.

Conclusions: The swim-up technique generates a sperm sample with better DNA integrity. Should DNA integrity correlate with better pregnancy rates in IUI and IVF, respectively, the swim-up may be the sperm processing technique of choice for these procedures.

Publication types

  • Comparative Study

MeSH terms

  • DNA / analysis
  • DNA / physiology*
  • Fertilization in Vitro / methods*
  • Humans
  • Male
  • Semen / chemistry
  • Semen / physiology*
  • Specimen Handling / methods*
  • Sperm Motility / physiology
  • Spermatozoa / chemistry
  • Spermatozoa / physiology*
  • Tolonium Chloride / chemistry*

Substances

  • Tolonium Chloride
  • DNA