Application of phosphoenolpyruvate into canine red blood cell cryopreservation with hydroxyethyl starch.
Phosphoenolpyruvate (PEP) is a phosphorylated glycolytic intermediate that can penetrate the RBC membrane and be metabolized to 2,3-DPG and ATP. In this study, we evaluated the effects of PEP treatment on canine red blood cells (RBCs) cryopreserved with 12.5% (w/v) HES. RBCs were incubated for 30, 60, and 90 min at 37 degrees C with PEP solution containing 60 mM mannitol, 30 mM sodium chloride, 25 mM glucose, 1 mM adenine and 50 mM PEP (340 m osm/kg), pH 6.0 and then cryopreserved in liquid nitrogen with 12.5% (w/v) HES for 2 weeks. 2,3-DPG and saline stabilities of the PEP treated groups were increased and osmotic fragility indices were significantly decreased compared to the untreated control group. There were no differences in 2,3-DPG levels within the PEP treated groups with different PEP incubation times. These results suggest that PEP treatment may be beneficial for the cryopreservation of canine RBCs with HES.
Kim
Heejaung
H
The United Graduate School of Veterinary Sciences, Yamaguchi University, Yoshida, Yamaguchi, Japan. b3646@yamaguchi-u.ac.jp
Itamoto
Kazuhito
K
Une
Satoshi
S
Nakaichi
Munekazu
M
Taura
Yasuho
Y
Sumida
Sajio
S
D019794:2,3-Diphosphoglycerate / Q000378:metabolism; D000818:Animals; D001793:Blood Preservation / Q000295:instrumentation* / Q000379:methods*; D015925:Cryopreservation / Q000295:instrumentation* / Q000379:methods*; D003451:Cryoprotective Agents / Q000378:metabolism / Q000494:pharmacology; D004285:Dogs; D004912:Erythrocytes / Q000166:cytology* / Q000378:metabolism; D006895:Hydroxyethyl Starch Derivatives / Q000494:pharmacology*; D009996:Osmotic Fragility; D010728:Phosphoenolpyruvate / Q000378:metabolism / Q000494:pharmacology*
D003451:Cryoprotective Agents; D006895:Hydroxyethyl Starch Derivatives; D019794:2,3-Diphosphoglycerate; D010728:Phosphoenolpyruvate