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Abstract

Volume 24, Issue 5 (September 2022) 24, 445–450; 10.4103/aja202197

MACS-annexin V cell sorting of semen samples with high TUNEL values decreases the concentration of cells with abnormal chromosomal content: a pilot study

Sahar El Fekih1,2, Nadia Gueganic1, Corinne Tous3, Habib Ben Ali2, Mounir Ajina4, Nathalie Douet-Guilbert1,3, Hortense Drapier3, Damien Beauvillard3, Frédéric Morel1,3, Aurore Perrin1,3

1 University of Brest, Inserm, UMR U1078, Faculty of Medicine and Health Sciences, Brest 29238, France
2 Laboratory of Human Cytogenetics, Molecular Genetics and Reproductive Biology, Farhat Hached University Hospital, Sousse 4031, Tunisia
3 Departement of Medical Genetics and Reproductive Biology, Brest University Regional Hospital, Brest 29609, France
4 Department of Reproductive Medicine, Farhat Hached Teaching Hospital, Sousse 4031, Tunisia

Correspondence: Dr. F Morel (frederic.morel@chu-brest.fr)

Date of Submission 07-Jul-2021 Date of Acceptance 28-Oct-2021 Date of Web Publication 31-Dec-2021

Abstract

We question whether, in men with an abnormal rate of sperm DNA fragmentation, the magnetic-activated cell sorting (MACS) could select spermatozoa with lower rates of DNA fragmentation as well as spermatozoa with unbalanced chromosome content. Cryopreserved spermatozoa from six males were separated into nonapoptotic and apoptotic populations. We determined the percentages of spermatozoa with (i) externalization of phosphatidylserine (EPS) by annexin V-Fluorescein isothiocyanate (FITC) labeling, (ii) DNA fragmentation by TdT-mediated-dUTP nick-end labeling (TUNEL), and (iii) numerical abnormalities for chromosomes X, Y, 13, 18, and 21 by fluorescence in situ hybridization (FISH), on the whole ejaculate and selected spermatozoa in the same patient. Compared to the nonapoptotic fraction, the apoptotic fraction statistically showed a higher number of spermatozoa with EPS, with DNA fragmentation, and with numerical chromosomal abnormalities. Compared to the whole ejaculate, we found a significant decrease in the percentage of spermatozoa with EPS and decrease tendencies of the DNA fragmentation rate and the sum of disomy levels in the nonapoptotic fraction. Conversely, we observed statistically significant higher rates of these three parameters in the apoptotic fraction. MACS may help to select spermatozoa with lower rates of DNA fragmentation and unbalanced chromosome content in men with abnormal rates of sperm DNA fragmentation.

Keywords: chromosomal abnormalities; DNA fragmentation; externalization of phosphatidylserine; magnetic-activated cell separation; spermatozoa

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