Clin Exp Reprod Med.  2019 Mar;46(1):8-13. 10.5653/cerm.2019.46.1.8.

Evaluation of normal morphology, DNA fragmentation, and hyaluronic acid binding ability of human spermatozoa after using four different commercial media for density gradient centrifugation

• Lee D
• Jee BC

• KMID: 2469601
• DOI: http://doi.org/10.5653/cerm.2019.46.1.8

Abstract

OBJECTIVE
Density gradient centrifugation (DGC) is frequently used to isolate high-motility fractions of spermatozoa. We compared the efficacy of four DGC media in terms of the percentage of morphologically normal spermatozoa, DNA fragmentation level, and hyaluronic acid (HA) binding ability.
METHODS
Thirty men with a total motile spermatozoa count >80 million participated. Semen samples were divided into four aliquots, which were processed using PureSperm, PureCeption, Sidney, and SpermGrad media, respectively. The DNA fragmentation level was measured using the Halosperm assay kit and HA binding ability was measured using the HBA assay kit.
RESULTS
The mean percentage of morphologically normal spermatozoa was significantly enhanced after DGC using all four media (10.3%, 9.9%, 9.8%, and 10.7%, respectively; p<0.05 for each when compared with 6.9% in raw semen). The DNA fragmentation level was significantly reduced after DGC using PureSperm, PureCeption, and SpermGrad media (6.0%, 6.5%, and 4.9%, respectively; p<0.05 for each when compared with 11.2% in raw semen), but not after DGC using Sidney media (8.5%, p>0.05). HA binding ability did not change after DGC using any of the four media.
CONCLUSION
The four media were equally effective for obtaining a sperm fraction with highly motile, morphologically normal sperm. PureSperm, PureCeption, and SpermGrad media were equally effective for acquiring a sperm fraction with less DNA fragmentation.

Keyword

Centrifugation, density gradient; DNA fragmentation; Hyaluronic acid; Spermatozoa

MeSH Terms

Centrifugation, Density Gradient*
DNA Fragmentation*
DNA*
Humans*
Hyaluronic Acid*
Male
Semen
Spermatozoa*
DNA
Hyaluronic Acid