Impact of cryopreservation agents on sperm quality, DNA fragmentation, and apoptotic markers in fertile and infertile males

Semen cryopreservation is a crucial technique for preserving male fertility, playing a vital role in assisted reproductive procedures by storing frozen semen samples for artificial insemination (AI) and intra-cytoplasmic sperm injection (ICSI) to enhance reproductive success rates. This study aims to identify the most effective cryopreservation methods and assess their impact on semen quality, particularly sperm DNA fragmentation. A total of 30 semen samples were categorized into fertile and infertile groups. DNA fragmentation analysis was conducted using the Sperm Chromatin Structure Assay (SCSA). Each sample was divided into three portions and frozen using different cryoprotectants: (egg-yolk + glycerol), (sucrose + glycerol), and (glycerol alone). After one month of storage, the samples were analyzed to determine the most effective medium. The findings revealed a decline in sperm motility post-freezing compared to fresh samples, along with a slight increase in morphological abnormalities. Additionally, there was a rise in sperm DNA fragmentation and an increase in apoptotic marker (Caspase-3) levels after the freezing process. The study concluded that cryopreservation and thawing caused some degree of sperm cell damage, with infertile samples being more adversely affected than fertile ones.