Effects of Cumulus Cell Extracellular Matrix–Assisted Sperm Preparation Protocols on In Vitro Sperm Quality

This study aimed to determine the effects of adding cumulus cell extracellular matrix (CCECM) to the culture medium used in sperm preparation techniques for assisted reproductive treatments (ART) on routine sperm parameters, fertilization capacity, and sperm DNA fragmentation, and to compare sperm separation protocols. Ejaculate samples from 30 normozoospermic male patients and cumulus–oocyte complexes collected from their spouses were included. Each semen sample was divided into five equal volumes and allocated to five groups: control, density gradient (DG), density gradient plus cumulus cell extracellular matrix (DG+K), swim-up (SU), and swim-up plus cumulus cell extracellular matrix (SU+K). The results showed that sperm concentration was markedly lower in all experimental groups than in the control group, with the decrease reaching statistical significance, particularly in the swim-up group. Motility was highest in the SU+K group and was also significantly increased in the DG+K and SU groups. In contrast, the TPMSS value decreased significantly in the SU and SU+K groups. No marked differences were observed among the groups in terms of morphology. Viability increased notably in the CCECM-supplemented groups, and this increase remained significant across all experimental groups compared with the control group. ARIC scores, reflecting acrosomal reaction capacity, were significantly higher in the DG+K and SU+K groups. Moreover, DFI values, indicating DNA damage, were significantly lower in these two groups, suggesting that CCECM supplementation may help preserve sperm DNA integrity. Adding CCECM to the culture medium used in sperm washing improved sperm quality and efficiency, particularly with the swim-up method. We believe this approach may be a feasible option for treating male infertility within assisted reproductive technologies (ART).