Carboxylated ε-Poly-L-Lysine Supplementation of the Freezing Extender Improves the Post-Thawing Boar Sperm Quality

SIMPLE SUMMARY: Frozen boar sperm is used on a very limited scale in pig artificial insemination owing to the low quality of post-thaw sperm. Cryoprotectant is usually used in boar sperm freezing extender, which is important for improving the post-thaw sperm quality. The carboxylated ε-poly-L-lysine, an efficient and non-toxic cryoprotectant, has been used as a food and cell preservative, as well as for tissue engineering and drug delivery in the biomedical applications. However, whether addition of carboxylated ε-poly-L-lysine to the freezing medium improves the post-thaw boar sperm quality or not is unknown. In this study, the addition of 0.25% carboxylated ε-poly-L-lysine to the freezing medium significantly improved the post-thaw boar sperm quality by protecting sperm mitochondrial function and antioxidant defense system. ABSTRACT: Frozen boar sperm is used on a minimal scale in consequence of the cryo-injuries induced by biochemical and physical modifications during the freezing and thawing processes. The present study investigates whether the addition of carboxylated ε-poly-L-lysine (CPLL) to the freezing medium could improve post-thaw boar sperm quality or not. Boar sperm was diluted with freezing medium contained different doses of carboxylated ε-poly-L-lysine (0, 0.125%, 0.25%, 0.5%, and 1%; v/v). The motility patterns, membrane integrity, acrosome integrity, mitochondrial membrane potential, NADH-CoQ activity, ATP level, malondialdehyde (MDA) level, and antioxidant defense system, as well as apoptosis in post-thaw boar sperm, were measured. It was observed that 0.25% CPLL treatment significantly improved the post-thaw boar sperm total motility, progressive motility, straight-linear velocity (VSL), curvilinear velocity (VCL), average path velocity (VAP), linearity (LIN), straightness (STR), membrane integrity, and acrosome integrity. Interestingly, the addition of CPLL also significantly increased the post-thaw sperm mitochondrial membrane potential, NADH-CoQ activity, and ATP level. Moreover, post-thaw boar sperm catalase (CAT) activity, glutathione peroxidase (GPx) activity, and superoxide dismutase (SOD) activity were increased with the addition of CPLL from 0.125% to 0.5% concentration levels. Furthermore, reduction of post-thaw sperm MDA level and apoptosis in 0.25% CPLL treatment was also observed. Those observations suggested that the addition of 0.25% CPLL to the freezing medium increased post-thaw boar sperm quality by protecting sperm mitochondrial function and antioxidant defense system. These findings provided novel insights that CPLL can be used as an efficient cryoprotectant to improve the post-thaw boar sperm quality during cryopreservation.