Subzero nonfreezing cryopresevation of rat hearts using antifreeze protein I and antifreeze protein III.
The purpose of the present study was to evaluate whether AFPs protect the heart from freezing and improve survival and viability in subzero cryopreservation. Hearts were subject to 5 preservation protocols; University of Wisconsin solution (UW) at 4 degrees C, UW at -1.3 degrees C without nucleation, UW at -1.3 degrees C with nucleation, UW AFP I (15 mg/cm(3)) at -1.3 degrees C with nucleation, and in UW AFP III (15 mg/cm(3)) at -1.3 degrees C with nucleation. Hearts were preserved for 24, 28, and 32 h, rewarmed and connected to the working isolated perfusion system. Data [heart rate (HR), coronary flow (CF), and developed pressure (dP)] was collected 30 and 60 min after reperfusion. Hearts preserved at -1.3 degrees C without AFPs froze, while hearts preserved with AFP did not freeze when nucleation was initiated and survived. Survival and dP of hearts preserved for 24h at -1.3 degrees C using AFP III was better than those preserved at 4 degrees C, (dP; 1.4 vs. 0.8, p<0.05). Four of six hearts and six of six hearts died when preserved at 4 degrees C for 28 and 32 h, respectively, all of the hearts that were preserved at -1.3 degrees C with or without AFPs survived after 28 h (n=18) and 32 h (n=18). CF was higher in UW -1.3 degrees C group without attempted nucleation than in AFP I and AFP III groups after 28 and 32 h (3.4 vs. 1.7, p<0.05, and 3.4 vs. 1.7, p<0.05, respectively). In conclusion, AFPs were found to protect the heart from freezing and improve survival and dP (AFP III) in prolonged subzero preservation.
Amir
Gabriel
G
Department of Cardiac Surgery, Sheba Medical Center, Tel Hashomer, Israel. gabiamir@stanford.edu
Horowitz
Liana
L
Rubinsky
Boris
B
Yousif
Basheer Sheick
BS
Lavee
Jacob
J
Smolinsky
Aram K
AK
D000241:Adenosine; D000493:Allopurinol; D000818:Animals; D021322:Antifreeze Proteins, Type I / Q000494:pharmacology*; D021343:Antifreeze Proteins, Type III / Q000494:pharmacology*; D002314:Cardioplegic Solutions; D003326:Coronary Circulation; D015925:Cryopreservation / Q000379:methods*; D005978:Glutathione; D006321:Heart; D006339:Heart Rate; D007328:Insulin; D008297:Male; D009926:Organ Preservation / Q000379:methods*; D019188:Organ Preservation Solutions; D011887:Raffinose; D051381:Rats; D017207:Rats, Sprague-Dawley; D013997:Time Factors
D021322:Antifreeze Proteins, Type I; D021343:Antifreeze Proteins, Type III; D002314:Cardioplegic Solutions; D007328:Insulin; D019188:Organ Preservation Solutions; C055206:University of Wisconsin-lactobionate solution; D000493:Allopurinol; D005978:Glutathione; D000241:Adenosine; D011887:Raffinose