OBJECTIVE: The study is to develop a glycerol-free extender using glucose for canine sperm cryopreservation,. METHODS: Tris [hydroxymethyl] aminomethane (TRIS) extender. Canine sperm were cooled to 4°C in TRIS containing 44.4 mM glucose for 100 min, and then cooled
at 4°C in TRIS with glucose concentrations of 0 mM, 44.4 mM, 100 mM, 200 mM, or 300 mM for 30 min followed by cryopreservation. After thawing at 37°C for 25 sec, sperm motility, viability, and morphological abnormalities were evaluated. In addition, 300 mM glucose-TRIS was compared
to TRIS extenders with a final concentration of 5% glycerol. Sperm phosphatidylserine (PS) translocation after freezing and thawing was assayed by flow cytometry using Annexin V-FITC apoptosis detection kit. RESULTS: Progressive motility and viability (42% and 41%, respectively) were
significantly higher in the 300mM group than the other groups with lower concentrations of glucose (P<0.05). PS translocation index was significantly lower in 300mM glucose-TRIS than those in extenders with glycerol (85 vs 93, P<0.05). CONCLUSION: These results indicate
that cryopreservation of canine sperm using glycerol-free 300 mM glucose-TRIS is feasible and yields more motile sperm with lower PS translocation compared with extenders containing glycerol.
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Keywords:
CANINE;
CRYOPRESERVATION;
GLUCOSE;
GLYCEROL-FREE TRIS;
PS TRANSLOCATION;
SPERMATOZOA
Document Type: Research Article
Publication date:
01 March 2014
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CryoLetters is a bimonthly international journal for low temperature sciences, including cryobiology, cryopreservation or vitrification of cells and tissues, chemical and physical aspects of freezing and drying, and studies involving ecology of cold environments, and cold adaptation
The journal publishes original research reports, authoritative reviews, technical developments and commissioned book reviews of studies of the effects produced by low temperatures on a wide variety of scientific and technical processes, or those involving low temperature techniques in the investigation of physical, chemical, biological and ecological problems.