title: Subzero nonfreezing cryopresevation of rat hearts using antifreeze protein I and antifreeze protein III.
abstract: 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.
authors.affiliation: Department of Cardiac Surgery, Sheba Medical Center, Tel Hashomer, Israel. gabiamir@stanford.edu
authors.forename: Gabriel
authors.initials: G
authors.lastname: Amir
authors.forename: Liana
authors.initials: L
authors.lastname: Horowitz
authors.forename: Boris
authors.initials: B
authors.lastname: Rubinsky
authors.forename: Basheer Sheick
authors.initials: BS
authors.lastname: Yousif
authors.forename: Jacob
authors.initials: J
authors.lastname: Lavee
authors.forename: Aram K
authors.initials: AK
authors.lastname: Smolinsky
mesh_terms: 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
chemical_list: 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