DOG SPERM CRYOPRESERVATION USING ONE STEP DILUTION WITH GLYCEROL-FREE TRIS EXTENDER

作者：Rahman Md Ataur; Park Sang Hyun; Yu Il Jeoung^*

来源：Cryo-Letters, 2016, 37(2): 137-141.

• canine sperm
• cryopreservation
• DNA integrity
• glycerol-free TRIS
• PS translocation

OBJECTIVE: The present study was to investigate a freezing method using one step-dilution with glycerol-free TRIS extender containing 172.2 mM glucose (GFTG). MATERIALS AND TMETHODS: The sperm pellet from selected ejaculates was resuspended in GFTG at 1 x 10(8) cells/mL. The semen was cooled for 10, 30, 50 or 70 min in GFTG at 4 degrees C and was frozen in LN2 vapor or in deep freezer (-80 degrees C, DF) for 20 min before plunge into LN2. Post-thaw sperm characteristics were examined. The phosphatidylserine (PS) translocation (Annexin V-FITC) and DNA integrity (TUNEL assay) were assessed using flow cytometry. RESULTS: Progressive motility and viability were significantly higher in 50 and 70 min groups than the other groups (P<0.05). PS translocation index was significantly lower in spermatozoa cooled for 50 or 70 min compared to 10 min (P<0.05). Freezing methods using LN2 vapor showed higher progressive motility than DF method (P<0.05), while viability and DNA fragmentation were not different between two freezing methods. CONCLUSION: Cryopreservation of canine sperm cooled for 50 or 70 min following one step dilution in GFTG yields more viable sperm with lower PS translocation and freezing method using LN2 vapor is more effective on progressive motility.

• 出版日期2016-4