Received February 21, 2024
Accepted August 25, 2024

Objective: To investigate the effect of astaxanthin supplementation in cryopreservation media on post-thawed sperm motility, viability, morphology, reactive oxygen species (ROS), and DNA fragmentation in two cryopreserved technique including vitrification and liquid nitrogen vapour freezing. Methods: Thirty normozoospermic semen samples were used in the study. Post-prepared semen samples were divided into 1) non-cryopreserved control, 2) and 3) vitrified without (V) and with astaxanthin 0.5 µM (V+ATX), 4) and 5) frozen in liquid nitrogen vapour without (L) and with astaxanthin 0.5 µM (L+ATX). Results: Sperm after cryopreservation using vitrification and liquid nitrogen vapour freezing was significantly decreased in motility, viability, and significantly increased ROS level but no change in sperm morphology and DNA fragmentation. The addition of astaxanthin in cryopreservation media significantly increases post-thawed motility in both vitrification (77.6 ± 8.9% vs 69.0 ± 9.5% in V+ATX and V) and vapour freezing (57.0 ± 13.3% vs 47.7 ± 14.6% in L+ATX and L), significantly increase sperm viability in vitrification (75.0 ± 11.9% vs 65.9 ± 11.1% in V+ATX and V), and significantly decrease ROS level in both vitrification (4.7 (2.6-8.3) RLU/sec/106 vs 10.6 (9.4-16.0) RLU/sec/106 in V+ATX and V) and vapour freezing (4.6 (3.3-10.5) RLU/sec/106 vs 10.3 (7.9-18.6) RLU/sec/106 in L+ATX and L). Astaxanthin supplementation in cryopreservation media does not affect sperm morphology and DNA fragmentation. Conclusion: Astaxanthin supplementation improved post-cryopreserved sperm motility, decreased ROS level in both vitrification and liquid nitrogen vapour freezing and improved sperm viability only in the vitrification technique.