title: Small molecule ice recrystallization inhibitors mitigate red blood cell lysis during freezing, transient warming and thawing.
abstract: During cryopreservation, ice recrystallization is a major cause of cellular damage. Conventional cryoprotectants such as dimethyl sulfoxide (DMSO) and glycerol function by a number of different mechanisms but do not mitigate or control ice recrystallization at concentrations utilized in cryopreservation procedures. In North America, cryopreservation of human red blood cells (RBCs) utilizes high concentrations of glycerol. RBC units frozen under these conditions must be subjected to a time-consuming deglycerolization process after thawing in order to remove the glycerol to <1% prior to transfusion thus limiting the use of frozen RBC units in emergency situations. We have identified several low molecular mass ice recrystallization inhibitors (IRIs) that are effective cryoprotectants for human RBCs, resulting in 70-80% intact RBCs using only 15% glycerol and slow freezing rates. These compounds are capable of reducing the average ice crystal size of extracellular ice relative to a 15% glycerol control validating the positive correlation between a reduction in ice crystal size and increased post-thaw recovery of RBCs. The most potent IRI from this study is also capable of protecting frozen RBCs against the large temperature fluctuations associated with transient warming.
authors.affiliation: Department of Chemistry, University of Ottawa, Ottawa, ON, K1N 6N5, Canada.
authors.forename: Jennie G
authors.initials: JG
authors.lastname: Briard
authors.affiliation: Department of Chemistry, University of Ottawa, Ottawa, ON, K1N 6N5, Canada.
authors.forename: Jessica S
authors.initials: JS
authors.lastname: Poisson
authors.affiliation: Canadian Blood Services, Centre of Innovation, 8249-114 Street NW, Edmonton, AB, T6G 2R8, Canada.
authors.forename: Tracey R
authors.initials: TR
authors.lastname: Turner
authors.affiliation: Department of Chemistry, University of Ottawa, Ottawa, ON, K1N 6N5, Canada.
authors.forename: Chantelle J
authors.initials: CJ
authors.lastname: Capicciotti
authors.affiliation: Canadian Blood Services, Centre of Innovation, 8249-114 Street NW, Edmonton, AB, T6G 2R8, Canada.
authors.forename: Jason P
authors.initials: JP
authors.lastname: Acker
authors.affiliation: Department of Chemistry, University of Ottawa, Ottawa, ON, K1N 6N5, Canada.
authors.forename: Robert N
authors.initials: RN
authors.lastname: Ben
mesh_terms: D001793:Blood Preservation / Q000379:methods*; D002470:Cell Survival / Q000187:drug effects; D003080:Cold Temperature; D015925:Cryopreservation / Q000379:methods*; D003451:Cryoprotective Agents / Q000737:chemistry / Q000494:pharmacology*; D003460:Crystallization; D004912:Erythrocytes / Q000187:drug effects* / Q000378:metabolism; D005615:Freezing; D005990:Glycerol / Q000494:pharmacology; D006461:Hemolysis / Q000187:drug effects*; D006801:Humans; D007053:Ice; D008856:Microscopy, Fluorescence / Q000379:methods; D018715:Microscopy, Video / Q000379:methods; D015394:Molecular Structure; D054852:Small Molecule Libraries / Q000737:chemistry / Q000494:pharmacology*
chemical_list: D003451:Cryoprotective Agents; D007053:Ice; D054852:Small Molecule Libraries; D005990:Glycerol