title: Canine sperm cryopreservation using glucose in glycerol-free Tris.
abstract: 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 degree C in TRIS containing 44.4 mM glucose for 100 min, and then cooled at 4 degree 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 degree 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).
  
  
  CONCLUSIONS
  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.
authors.affiliation: Department of Theriogenology and Reproductive Biotechnologies, College of Veterinary Medicine and Bio-safety Research Institute, Chonbuk National University, Jeonju, Republic of Korea. iyu@jbnu.ac.kr.
authors.forename: Il-Jeoung
authors.initials: IJ
authors.lastname: Yu
mesh_terms: D000818:Animals; D017304:Annexin A5; D002470:Cell Survival / Q000187:drug effects; D015925:Cryopreservation; D003451:Cryoprotective Agents / Q000494:pharmacology*; D004285:Dogs; D005434:Flow Cytometry; D005615:Freezing; D005947:Glucose / Q000494:pharmacology*; D005990:Glycerol / Q000494:pharmacology; D008297:Male; D010718:Phosphatidylserines / Q000378:metabolism; D012662:Semen Preservation / Q000379:methods*; D013076:Sperm Count; D013081:Sperm Motility / Q000187:drug effects; D013094:Spermatozoa / Q000187:drug effects* / Q000502:physiology / Q000648:ultrastructure; D014325:Tromethamine / Q000494:pharmacology*
chemical_list: D017304:Annexin A5; D003451:Cryoprotective Agents; D010718:Phosphatidylserines; D014325:Tromethamine; D005947:Glucose; D005990:Glycerol