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Updated: Aug 29, 2026

Flow Cytometric Analysis of Biomarkers for Detecting Human Sperm Functional Defects
Published on: April 21, 2022
Effects of age on DNA double-strand breaks and apoptosis in human sperm
Narendra P Singh1, Charles H Muller, Richard E Berger
1Department of Bioengineering, University of Washington, Seattle, Washington 98195-7962, USA. narendra@u.washington.edu
Objective:
This study was designed to explore the relationship between men's age and DNA damage and apoptosis in human spermatozoa.
Design:
Semen samples were collected from men between the ages of 20 and 57 years. Sperm DNA double-strand breaks were assessed using the neutral microgel electrophoresis (comet) assay, and apoptosis was estimated using the DNA diffusion assay.
Setting:
Academic medical center.
Patient(S):
Sixty-six men aged 20 to 57 years were recruited from infertility laboratory and general populations and consented to donate a semen sample. Recruitment was determined by time and day of analysis; the only exclusions were for azoospermia, prostatitis, or prior cancer therapy.
Intervention(S):
None.
Main Outcome Measure(S):
DNA damage and apoptosis in human sperm.
Result(S):
Age correlated with an increasing percentage of sperm with highly damaged DNA (range: 0-83%) and tended to inversely correlate with percentage of apoptotic sperm (range: 0.3%-23%). For example, percentage of sperm with highly damaged DNA, comet extent, DNA break number, and other comet measures was statistically significantly higher in men aged 36-57 years than in those aged 20-35 years, but percentage apoptosis was statistically significantly lower in the older group. Semen analysis showed percentage motility to be significantly higher in younger age groups.
Conclusion(S):
This study clearly demonstrates an increase in sperm double-stranded DNA breaks with age. Our findings also suggest for the first time an age-related decrease in human sperm apoptosis. These novel findings may indicate deterioration of healthy sperm cell selection process with age.
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