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Dynamic assessment of human sperm DNA damage II: the effect of sperm concentration adjustment during processing

  • Gamete Biology
  • Published:
Journal of Assisted Reproduction and Genetics Aims and scope Submit manuscript

Abstract

Purpose

To evaluate the effect of sperm concentration adjustment in human ejaculates on the sperm DNA quality and longevity.

Methods

Semen samples were obtained from 30 donors with a normal spermiogram. Following centrifugation, the sperm pellet was resuspended in PBS, and the sperm concentration adjusted to 200, 100, 50, 25, 12, and 6 × 106/mL. Each set of samples was incubated at 37 °C for 24 h, and the sperm DNA damage was assessed using the chromatin-dispersion test following 0 h, 2 h, 6 h, and 24 h of incubation.

Results

Sperm DNA fragmentation (SDF) did not differ between the selected experimental conditions at T0; however, Kaplan–Meier estimates for survival showed significant differences with respect to the dilution and time (all P values were smaller than .001). DNA fragmentation in semen samples adjusted to 200 × 106/mL was approximately 3.3 times higher when compared to samples containing 25 × 106/mL and 3.9 higher in comparison with samples adjusted to 12 × 106/mL following 2 h of in vitro incubation. Although there was evidence of individual variation in SDF during the incubation period, the general finding was that lower sperm concentrations resulted in a slower rate of DNA fragmentation.

Conclusions

Incubation of spermatozoa for ART purposes should be done following a concentration adjustment below 25 × 106/mL in order to avoid a higher susceptibility of the sperm DNA molecule towards fragmentation.

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Funding

This study was funded by the Spanish Ministry of Economy, Industry and Competitiveness—Programa Retos RTC-2016-4733-1 and by the Slovak Research and Development Agency Grant no APVV-15-0544. The funding bodies had no involvement in the study.

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Authors and Affiliations

  1. Department of Animal Physiology, Faculty of Biotechnology and Food Sciences, Slovak University of Agriculture in Nitra, Tr. A. Hlinku 2, 94976, Nitra, Slovakia

    Eva Tvrdá & Michal Ďuračka

  2. Unit of Genetics, Department of Biology, Universidad Autónoma de Madrid, Madrid, Spain

    Francisca Arroyo, Carmen López-Fernández & Jaime Gosálvez

Authors
  1. Eva Tvrdá
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  2. Francisca Arroyo
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  3. Michal Ďuračka
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  4. Carmen López-Fernández
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  5. Jaime Gosálvez
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Correspondence to Eva Tvrdá.

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All procedures performed in the study were in accordance with the ethical standards of the institutional research committee, with the 1964 Helsinki declaration and its later amendments or comparable ethical standards.

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Tvrdá, E., Arroyo, F., Ďuračka, M. et al. Dynamic assessment of human sperm DNA damage II: the effect of sperm concentration adjustment during processing. J Assist Reprod Genet 36, 799–807 (2019). https://doi.org/10.1007/s10815-019-01423-y

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  • DOI: https://doi.org/10.1007/s10815-019-01423-y

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