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Canine sperm vitrification with sucrose: effect on sperm function
Version of Record online: 24 MAR 2011
© 2011 Blackwell Verlag GmbH
Volume 43, Issue 4, pages 233–241, August 2011
How to Cite
Sánchez, R., Risopatrón, J., Schulz, M., Villegas, J., Isachenko, V., Kreinberg, R. and Isachenko, E. (2011), Canine sperm vitrification with sucrose: effect on sperm function. Andrologia, 43: 233–241. doi: 10.1111/j.1439-0272.2010.01054.x
- Issue online: 5 JUL 2011
- Version of Record online: 24 MAR 2011
- Accepted: December 7, 2009
- Canine sperm;
- sperm function;
The ability of sucrose to protect spermatozoa against mitochondrial damage, artificial acrosome reaction and DNA fragmentation during ultra-rapid cryopreservation in canine sperm was investigated. Swim-up selected spermatozoa of second-fraction semen were vitrified with different concentrations of sucrose (0.1, 0.25 and 0.4 m) in proportion 1 : 1 v/v with HTF–BSA 1%. From each group, 30-μl suspensions of cells were dropped directly into liquid nitrogen and stored for at least 24 h. Cells were thawed by submerging the spheres in HTF with 1% BSA at 37 °C. The number of progressively motile spermatozoa was significantly higher in the sucrose 0.25 m + HTF–BSA 1% (42.5 ± 2.3%, P < 0.01) than in HTF only (1.66 ± 0.3%). The same combination of sucrose 0.25 m + HTF–BSA 1% (42.7 ± 1.5%) had a stronger cryoprotective effect on the integrity of mitochondrial membrane potential (P < 0.05) and decreased the DNA fragmentation (2.8 ± 0.5%) as compared with HTF only (1.93 ± 0.6% and 5.6 ± 0.6% respectively). With respect to acrosome-reacted spermatozoa, no significant difference was found between the groups investigated (P > 0.05). It is concluded that sucrose, a nonpermeable cryoprotectant, can effectively preserve important physiological parameters such as mitochondrial membrane potential and DNA integrity during ultra-rapid cryopreservation.