Impact of Microporous Membrane-Based Sperm Selection on DNA Fragmentation Index Assessed by Sperm Chromatin Dispersion (CANfrag Kit) Assay

Abstract

Sperm DNA fragmentation (SDF) is a crucial indicator of fertility potential of males and is poorly assessed through traditional semen parameters. Although sperm processing enhances motility and morphology, the effects of such manipulations on sperm DNA integrity need further elucidation. Microporous membrane-assisted sperm separation provides a gentle technique that does not involve centrifugation in order to isolate functionally competent sperm cells. The primary goal of the study is to estimate the effect of microporous membrane-assisted sperm isolation (CANezy) on DNA fragmentation index (DFI), progressive motility, and morphology using the sperm chromatin dispersion (CANfrag) Kit test. Fifty semen samples from males referred for fertility evaluation were processed with the use of CANezy microporous membrane device (Candore Bioscience). Only samples that met WHO criteria (volume ≥1.0 mL, concentration ≥5 million/mL, complete liquefaction) were used. Sperm parameters (concentration, motility, morphology) and DFI were analyzed before and after microporous membrane separation. CANfrag kit test was used to assess sperm DNA fragmentation. Paired statistical analysis was carried out to compare sperm parameters before and after sperm isolation. Microporous membrane separation enhanced all semen parameters. Progressive motility mean value increased from 44.10% ± 9.85% to 89.66% ± 3.24% (P < 0.001). The normal morphology increased from 12.08% ± 4.89% to 24.14% ± 7.68% (P < 0.001). The most remarkable improvement was the drop in the DNA fragmentation index from 25.92% ± 8.74% in raw semen samples to 7.64% ± 3.92% after separation (P < 0.001), which is equivalent to a 70.5% decrease. The sperm concentration reduced from 68.08 ± 28.54 million/mL to 33.90 ± 17.82 million/mL because of the selective enrichment of good quality sperms. Micro porous membrane-based sperm separation with the CANezy device leads to a significant reduction of sperm DNA fragmentation while at the same time improving the motility and morphology. It is a mild procedure that selectively enriches functionally competent spermatozoa with intact chromatin integrity. Clinical advantages for assisted reproductive technologies, especially when there are high baseline DNA fragmentation levels, are expected.