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DNA fragmentation in human sperm after magnetic-activated cell sorting

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

Abstract

Purpose

As fertilization with unselected apoptotic spermatozoa may contribute to failures in assisted reproductive techniques, it has become essential to remove this type of sperm in order to increase the success rates. Magnetic-activated cell sorting (MACS) is a sperm preparation technique that isolates non-apoptotic spermatozoa based on the expression of phosphatidylserine in the membrane of apoptotic sperm. Therefore, we aimed to evaluate whether there was a significant decrease in sperm DNA fragmentation (sDNAfrag) and verify which protocol was the most efficient.

Methods

Hundred semen samples were allocated into five distinct groups and processed according to a combination of MACS with density gradient centrifugation (DGC) and swim-up (SU) techniques. Sperm DNA fragmentation was evaluated by terminal deoxynucleotidyl transferase dUTP nick-end labeling (TUNEL) assay.

Results

Groups DGC-SU (73.4 %), DGC-MACS-SU (78.9 %), DGC-SU-MACS (53.8 %) and MACS-SU (73.5 %) presented a significant decrease in sDNAfrag but the highest reduction rate was obtained with MACS-DGC-SU (83.3 %). The later was also negatively correlated with sperm vitality, membrane integrity and progressive motility. Additionally, teratozoospermic patients presented a tendency to have lower sDNAfrag reduction rates than asthenozoospermic and asthenoteratozoospermic patients.

Conclusions

Based on the results, MACS showed potential to optimize the sDNAfrag reduction rate, when applied to raw semen, before DGC and SU, especially in samples with low values of progressive motility, vitality and hypoosmotic swelling test.

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Acknowledgments

We would like to acknowledge Ângela Alves, Technical assistant for teaching and research (ICBAS-UP) in laboratorial assistance, Claúdia Osório, Bsc, Biologist (CGR-ABarros) in spermiogram analysis assistance and Gabriela Rodrigues, PhD, Assistant Professor (University of Lisbon) for reviewing the manuscript.

Multidisciplinary Unit for Biomedical Research (UMIB) is funded by National Funds through FCT-Foundation for Science and Technology, under the Pest-OE/SAU/UI0215/2014.

Conflict of interest

The authors declare that they have no conflict of interest.

Author information

Authors and Affiliations

  1. Department of Microscopy, Laboratory of Cell Biology, Institute of Biomedical Sciences Abel Salazar (ICBAS), University of Porto (UP), Multidisciplinary Unit for Biomedical Research (UMIB), Rua Jorge Viterbo Ferreira, 228, 4050-313, Porto, Portugal

    Sara Bucar, Mário Sousa & Rosália Sá

  2. Department of Animal Biology, Faculty of Sciences, University of Lisbon, Campo Grande, 1749-016, Lisbon, Portugal

    Sara Bucar

  3. Centre for Reproductive Genetics Prof. Alberto Barros (CGR-ABarros), Av. do Bessa, 240, 1° Dto. Frente, 4100-009, Porto, Portugal

    Ana Gonçalves & Alberto Barros

  4. Department of Microscopy, Laboratory of Histology and Embryology, ICBAS-UP, Rua Jorge Viterbo Ferreira, 228, 4050-313, Porto, Portugal

    Eduardo Rocha

  5. Department of Genetics, Faculty of Medicine, UP, Alameda Prof. Hernâni Monteiro, 4200-319, Porto, Portugal

    Alberto Barros

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  1. Sara Bucar
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  2. Ana Gonçalves
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  4. Alberto Barros
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  6. Rosália Sá
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Corresponding author

Correspondence to Rosália Sá.

Additional information

Capsule Semen was processed with MACS and classic sperm preparation techniques. The highest reduction of sDNAfrag was obtained with MACS followed by DGC and SU and in asthenozoospermic patients. Sperm progressive motility, vitality and membrane integrity were negatively correlated with sDNAfrag.

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Bucar, S., Gonçalves, A., Rocha, E. et al. DNA fragmentation in human sperm after magnetic-activated cell sorting. J Assist Reprod Genet 32, 147–154 (2015). https://doi.org/10.1007/s10815-014-0370-5

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  • DOI: https://doi.org/10.1007/s10815-014-0370-5

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