Sperm quality after density gradient centrifugation with three commercially available media: a controlled trial

Helena Malvezzi, Rakesh Sharma, Ashok Agarwal, Adel M Abuzenadah, Muhammad Abu-Elmagd, Helena Malvezzi, Rakesh Sharma, Ashok Agarwal, Adel M Abuzenadah, Muhammad Abu-Elmagd

Abstract

Background: Density gradient is the preferred technique for sperm processing for ART. However, no study has examined sperm quality using different processing media simultaneously and under identical conditions. Therefore, we evaluated semen quality following sperm preparation by three commonly used commercially available density gradient media in a well-designed controlled trial.

Methods: We obtained semen samples from 20 healthy volunteers. Percent motility, total motile sperm (TMS), % recovery and DNA damage were assessed before and after separation in three different sperm density gradient media-PureCeption, ISolate and SpermGrad-125.

Results: Percent motility was higher in the ISolate (81.4% ± 6.6%) and SpermGrad-125 samples (85.7% ± 8.0%) (P < 0.0001) than in the PureCeption samples (62.5% ± 13.2%) (P = 0.07). TMS was higher in the PureCeption(TM) and ISolate samples (14.2% ± 15.9% and 15.8% ± 18.2%) than in those prepared with SpermGrad-125 (10.6% ± 19.7%) (P < 0.0001). Percent recovery was significantly higher in the PureCeption(TM) and ISolate samples (45.3% and 48.9%) than in the SpermGrad-125(TM) samples (30.8%) (P < 0.01). DNA fragmentation was comparable across the three gradients (PureCeption = 8.8% ± 4.7%; ISolate = 7.2 ± 5.2% and SpermGrad-125 = 11.2% ± 7.4%).

Conclusions: Three different density gradient processing media PureCeption, ISolate, and SpermGrad-125 were examined for their effects on sperm quality. Sperm processed by ISolate and Sperm Grad 125 had better motility and TMS after processing. The extent of DNA damage was comparable in all three gradients.

Figures

Figure 1
Figure 1
Schematics of TUNEL assay for measurement of DNA damage in spermatozoa.
Figure 2
Figure 2
Schematics of sperm preparation by density gradient separation.
Figure 3
Figure 3
Schematics of the DNA damage by the TUNEL assay. (1) Labeling of DNA strand breaks with TdT, which catalyzes the polymerization of labeled nucleotides to free 3′-OH DNA ends in a template-independent manner (TUNEL reaction) and (2) Fluorescein isothiocynate (FITC)-dUTP is incorporated into nucleotide polymers, and it can be directly detected and quantified by flow cytometry.

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