Cryo letters · 2014
Cryoprotectant-free freezing of the goat epididymal sperm.
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Katanbafzadeh H, Barati F, Tabandeh M
Аннотация
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.
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.