RESUMEN
A cryoprotectant-free method was successfully used for rapid freezing of goat epididymal spermatozoa. Lowering sperm volume may increase the temperature exchange rate and improve the freezing output of spermatozoa. The aim of this study was to compare two different packaging types [0.25 ml French straws (FS) and 96-well immune plate (WIP)] for rapid freezing of goat epididymal spermatozoa. Eleven pairs of the goat testes were transferred to the laboratory; cauda epididymidides were dissected and sliced in TRIS-BSA solution for 15 min and temperature 33-35 °C. Sperm concentration was adjusted to 20 × 10(6) ml(-1), and the suspension was subjected to rapid freezing within FS or WIP. The volume of spermatozoa in WIP method was set at 25 µl. Sperm motility, viability and abnormalities, and sperm DNA integrity were compared between two devices. The results showed similar effectiveness of WIP and FS on post-thaw sperm parameters. In conclusion, for cryoprotectant-free rapid freezing of goat epididymal spermatozoa, it is recommended to use WIP instead of French 0.25 ml straws.
Asunto(s)
Criopreservación/métodos , Preservación de Semen/métodos , Espermatozoides/fisiología , Animales , Fragmentación del ADN , Cabras , Masculino , Preservación de Semen/instrumentación , Motilidad EspermáticaRESUMEN
BACKGROUND: Cryoprotectant free approach successfully removed the impact of physical and chemical damages in preserving human sperm in a vitrification protocol. There is no any report on this technology in farm animal sperm freezing. OBJECTIVE: The aim of the present study was to find the efficacy of cholesterol-loaded cyclodextrin (CLC; 1 mg per 60 million) and sucrose (0.1 and 0.2 M) on freezing of the goat epididymal sperm. METHODS: Caudal epididymides (n=5 pairs) were dissected, incised and incubated in the Tris-BSA solution for 15 min, followed by swim-up at room temperature. Sperm was loaded in 0.25 mL French straws and cooled on nitrogen vapor for 3 min then immersed in liquid nitrogen and remained for 48 h. Then the straws thawed by immersing in 37 degree C waterbath for 30 sec and analyzed. RESULT: The results showed the impact of freezing on the goat epididymal sperm motility, viability and DNA fragmentation that were improved by incorporation of CLC and sucrose (0.2 M). CONCLUSIONS: In conclusion, the goat epididymal sperm was frozen in a cryoprotectant-free freezing model. CLC and 0.4 M sucrose protected the goat epididymal sperm against freezing-induced damages.