A new system of sperm cryopreservation: evaluation of survival, motility, DNA oxidation, and mitochondrial activity

Background Sperm vitrification (V) is a method for cryopreservation, without the use of conventional cryoprotectants, by plunging the sperm suspension directly into liquid nitrogen (LN25). Objective This study aimed to compare the new system of V with conventional freezing (CF) protocol using fresh...

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Bibliographic Details
Published in:Andrology (Oxford) 2019-05, Vol.7 (3), p.293-301
Main Authors: Pabón, D., Meseguer, M., Sevillano, G., Cobo, A., Romero, J. L., Remohí, J., de los Santos, M. J.
Format: Article
Language:English
Subjects:
Cell Survival
Cohort Studies
Cryopreservation
Cryopreservation - methods
cryoprotectants‐free
Deoxyribonucleic acid
DNA
DNA - metabolism
Freezing
human spermatozoa
Humans
Mitochondria - metabolism
Motility
Oxidation
Oxidation-Reduction
Prospective Studies
Semen Preservation - methods
Sperm
Sperm Motility
ultrarapid freezing
vitrification
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Summary:Background Sperm vitrification (V) is a method for cryopreservation, without the use of conventional cryoprotectants, by plunging the sperm suspension directly into liquid nitrogen (LN25). Objective This study aimed to compare the new system of V with conventional freezing (CF) protocol using fresh spermatozoa as reference (C). Material and methods Prospective cohort study. A total of 47 sperm samples from men attending the infertility clinic at Instituto Valenciano de Infertilidad Valencia. The sperm V solution was 0.3 M trehalose–sucrose and plunged directly in liquid nitrogen in microdroplets of 5–10 lL, using a new system collector of V. Sperm viability indicators such as sperm motility, vitality rates, mitochondrial function, and sperm DNA oxidation were assessed before and after cryopreservation. Sperm motility and vitality analysis were performed according to published guidelines of the World Health Organization (WHO, 2010). Mitochondrial function was evaluated using JC‐1 (fluorescent cationic dye, 5,50,6,60‐tetrachloro‐1‐10,3,30‐tetraethyl‐benzamidazolocarbocyanin iodide). Sperm DNA oxidation was determined using a fluorescent assay (Oxy‐DNA test) for the detection of 8‐oxoguanine. The evaluation was carried out before and after cryopreservation using flow cytometry. Statistical analysis was performed using ANOVA and chi‐square test, and p 
ISSN:2047-2919
2047-2927
DOI:10.1111/andr.12607