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Evaluation of conventional semen parameters, intracellular reactive oxygen species, DNA fragmentation and dysfunction of mitochondrial membrane potential after semen preparation techniques: a flow cytometric study

  • Reproductive Medicine
  • Published:
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Abstract

Purpose

The aim of this study was to evaluate the conventional sperm parameters, level of intracellular reactive oxygen species (ROS), DNA fragmentation (DF) and dysfunction of mitochondrial membrane potential (MMP) after semen preparation techniques with flow cytometry.

Methods

Semen samples were obtained from 28 men with normal semen analysis according to WHO (world health organization). Each was divided into three equal parts for processing with routine techniques: conventional swim up (CSW), direct swim up (DSW) and density gradient centrifugation (DGC). The conventional sperm parameters were evaluated with computer-assisted sperm analyzer (CASA) and the level of intracellular ROS, dysfunction of MMP and DF were determined with flow cytometry procedure.

Results

Conventional sperm parameters such as motility, progressive motility and normal morphology increased after sperm processing by CSW and DGC compared to DSW. A significant increase in intracellular H2O2 (p < 0.05) was demonstrated in the CSW versus DSW technique, while processed sperm by the DSW procedure showed a significant increase in the percentage of dysfunction of MMP and intracellular O •−2 (p < 0.05) when compared with CSW and DGC techniques. Additionally, a high mean of DF (p < 0.05) was observed in the DGC technique as compared to CSW.

Conclusion

Data from flow cytometry study demonstrated that intracellular H2O2 and DF increased after CSW and DGC processing techniques, respectively, whereas the level of intracellular O •−2 and dysfunction of MMP only increased after the DSW processing technique.

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Acknowledgments

The authors thank Dr. Bita Ebrahimi and Mr Fazel Samani for their assistance and helpful comments during this project.

Conflict of interest

None.

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Authors and Affiliations

  1. Department of Embryology at Reproductive Biomedicine Research Center, Royan Institute for Reproductive Biomedicine, ACECR, P. O. 19395-4644, Tehran, Iran

    Leila Rashki Ghaleno, Mojtaba Rezazadeh Valojerdi & Mina Sharbatoghli

  2. Department of Developmental Biology, University of Science and Culture, ACECR, Tehran, Iran

    Leila Rashki Ghaleno

  3. Department of Anatomy, Faculty of Medical Science, Tarbiat Modares University, P. O. 14115-111, Tehran, Iran

    Mojtaba Rezazadeh Valojerdi

  4. Department of Stem Cells and Developmental Biology at Cell Science Research Center, Royan Institute for Stem Cell Biology and Technology , ACECR, Tehran, Iran

    Ehsan Janzamin

  5. Department of Epidemiology and Reproductive Health at Reproductive Epidemiology Research Center, Royan Institute for Reproductive Biomedicine, ACECR, Tehran, Iran

    Mohammad Chehrazi

  6. Department of Andrology at Reproductive Biomedicine Research Center, Royan Institute for Reproductive Biomedicine, ACECR, Tehran, Iran

    Reza Salman Yazdi

Authors
  1. Leila Rashki Ghaleno
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  2. Mojtaba Rezazadeh Valojerdi
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Correspondence to Mojtaba Rezazadeh Valojerdi.

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Ghaleno, L.R., Valojerdi, M.R., Janzamin, E. et al. Evaluation of conventional semen parameters, intracellular reactive oxygen species, DNA fragmentation and dysfunction of mitochondrial membrane potential after semen preparation techniques: a flow cytometric study. Arch Gynecol Obstet 289, 173–180 (2014). https://doi.org/10.1007/s00404-013-2946-1

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  • DOI: https://doi.org/10.1007/s00404-013-2946-1

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