Related Experiment Video
Updated: May 28, 2026
Rapid and Efficient Spatiotemporal Monitoring of Normal and Aberrant Cytosine Methylation within Intact Zebrafish Embryos
Published on: August 18, 2022
Repeated assessment by high-throughput assay demonstrates that sperm DNA methylation levels are highly reproducible
Victoria K Cortessis1, Kimberly Siegmund, Sahar Houshdaran
1Department of Preventive Medicine, Keck School of Medicine, University of Southern California, Los Angeles, California 90033, USA. cortessi@usc.edu
Objective:
To assess reliability of a high-throughput assay of sperm DNA methylation.
Design:
Observational study comparing DNA methylation of sperm isolated from 3 divided and 12 longitudinally collected semen samples.
Setting:
Academic medical center.
Patient(S):
One man undergoing screening semen analysis during evaluation of an infertile couple and 2 healthy fertile male volunteers.
Intervention(S):
Spermatozoa were separated from seminal plasma and somatic cells using gradient separation. DNA was extracted from spermatozoa, and DNA methylation was assessed at 1,505 DNA sequence-specific sites.
Main Outcome Measure(S):
Repeatability of sperm DNA methylation measures, estimated by correlation coefficients.
Result(S):
DNA methylation levels were highly correlated within matched sets of divided samples (all r ≥ 0.97) and longitudinal samples (average r = 0.97).
Conclusion(S):
The described methodology reliably assessed methylation of sperm DNA at large numbers of sites. Methylation profiles were consistent over time. High-throughput assessment of sperm DNA methylation is a promising tool for studying the role of epigenetic state in male fertility.
