DNA fragmentation in concert with the simultaneous assessment of cell viability in a subfertile population: establishing thresholds of normality both before and after density gradient centrifugation.
Adult
Cell Survival
/ genetics
Centrifugation, Density Gradient
Chromatin
/ genetics
DNA Damage
/ genetics
DNA Fragmentation
Female
Fertility
/ genetics
Fertilization in Vitro
Humans
Infertility, Male
/ genetics
Male
Middle Aged
Pregnancy
Reproductive Techniques, Assisted
Semen
/ chemistry
Semen Analysis
Spermatozoa
/ chemistry
Density gradient centrifugation
Neat semen
TUNEL assay
Threshold values
Total sperm DNA fragmentation
Vital sperm DNA fragmentation
Journal
Journal of assisted reproduction and genetics
ISSN: 1573-7330
Titre abrégé: J Assist Reprod Genet
Pays: Netherlands
ID NLM: 9206495
Informations de publication
Date de publication:
Jul 2019
Jul 2019
Historique:
received:
25
01
2019
accepted:
03
05
2019
pubmed:
16
5
2019
medline:
18
12
2019
entrez:
16
5
2019
Statut:
ppublish
Résumé
TUNEL assay is the most common, direct test for sperm chromatin integrity assessment. But, lack of standardized protocols makes interlaboratory comparisons impossible. Consequently, clinical thresholds to predict the chance of a clinical pregnancy also vary with the technique adopted. This prospective study was undertaken to assess the incidence of sperm DNA fragmentation in a subfertile population and to establish threshold values of normality as compared to a fertile cohort, both before and after density gradient centrifugation in the total and vital fractions. Men presenting at a university hospital setup for infertility treatment. DNA damage via TUNEL assay was validated on fresh semen samples, as conventional semen parameters, to reduce variability of results. Total DNA fragmentation in the neat semen was significantly higher in the subfertile group, but the vital fraction was not significantly different between the two cohorts. After gradient centrifugation, DNA fragmentation increased significantly in the total fraction of the subfertile group but decreased significantly in the vital fraction. In the fertile cohort, there was a non-significant increase in total fragmentation and in the vital fraction the trend was unclear. Estimating total and vital sperm DNA fragmentation, after density gradient centrifugation, increased both the sensitivity and the specificity, thereby lowering the number of false negatives and false positives encountered. These findings provide opportunities to investigate the significance of the total and the vital fractions after different assisted reproductive technologies.
Identifiants
pubmed: 31089933
doi: 10.1007/s10815-019-01476-z
pii: 10.1007/s10815-019-01476-z
pmc: PMC6642232
doi:
Substances chimiques
Chromatin
0
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Pagination
1413-1421Subventions
Organisme : Fonds Wetenschappelijk Onderzoek
ID : T007016N
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