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Abstract

Volume 12, Issue 5 (September 2010) 12, 753–759; 10.1038/aja.2010.46

Preparation and incubation conditions affect the DNA integrity of ejaculated human spermatozoa

Rieko Matsuura, Takumi Takeuchi, Atsumi Yoshida

The Reproduction Center, Kiba Park Clinic, Tokyo 135-0042, Japan

Correspondence: Dr Takumi Takeuchi, ttakeuch@me.com

Received 9 December 2009; Revised 10 March 2010; Accepted 10 May 2010; Published online 21 June 2010.

Abstract

Appropriate semen processing and assessment are critical for successful infertility treatment. We investigated whether laboratory procedures including semen preparation and incubation affect sperm DNA integrity. A total of 153 infertile men were involved. Conventional semen parameters and sperm chromatin structure assay (SCSA) parameters, that is, DNA fragmentation index (%DFI) and high DNA stainability (%HDS), were assessed on the fresh ejaculated semen samples, which were treated and incubated under different conditions. Negative correlations were identified between the %DFI and sperm concentration, motility, progressive motility and morphology. A lower percentage of DFI was detected in spermatozoa when density gradient centrifugation (DGC) was followed by swim-up treatment in comparison with DGC alone (P < 0.01). Although the %DFI increased in a time-dependent manner with incubation both at room temperature (RT) and at 37°C in air, the %DFI after 24 h at RT was significantly lower than that at 37°C (P < 0.05). Incubation with 5% CO2 was effective in maintaining sperm motility (P < 0.01); however, it induced further elevation of %DFI (P < 0.001). Thus, sperm DNA damage was associated with longer incubation periods. Interestingly, common culture conditions, such as maintaining pH and temperature, compromised the sperm DNA integrity.


Keywords:

density gradient centrifugation; DNA damage; male infertility; sperm chromatin structure assay; spermatozoa

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