Flow Cytometric Chromosomal Sex Sorting of Stallion Spermatozoa Induces Oxidative Stress on Mitochondria and Genomic DNA

• Published in: Reproduction in domestic animals 2016-02, Vol.51 (1), p.18-25
• Main Authors: Balao da Silva, CM, Ortega-Ferrusola, C, Morrell, JM, Rodriguez Martínez, H, Peña, FJ
• Format: Article
• Language: English
• Subjects: Animals; Annan veterinärmedicin; Cell Separation - methods; Cell Separation - veterinary; DNA Damage; DNA Fragmentation; DNA, Mitochondrial - genetics; Flow Cytometry - veterinary; Horses - genetics; Male; Membrane Potential, Mitochondrial; Other Veterinary Science; Oxidative Stress - genetics; Semen - physiology; Sex Preselection - methods; Sex Preselection - veterinary; Sperm Motility; Spermatozoa - physiology
• ISSN: 0936-6768; 1439-0531; 1439-0531
• DOI: 10.1111/rda.12640

To date, the only repeatable method to select spermatozoa for chromosomal sex is the Beltsville sorting technology using flow cytometry. Improvement of this technology in the equine species requires increasing awareness of the modifications that the sorting procedure induces on sperm intactness. Oxidative stress is regarded as the major damaging phenomenon, and increasing evidence regards handling of spermatozoa – including sex sorting – as basic ground for oxidative damage. The aim of this study was to disclose whether the flow cytometric sorting procedure increases the production of reactive oxygen species (ROS), and to identify if ROS production relates to DNA damage in sorted spermatozoa using specific flow cytometry‐based assays. After sorting, oxidative stress increased from 26% to 33% in pre‐ and post‐incubation controls, to 46% after sex sorting (p