title: Influence of intracellular trehalose concentration and pre-freeze cell volume on the cryosurvival of rapidly frozen human erythrocytes.
abstract: Significant interest exists in the application of trehalose, which has low permeability to the phospholipid bilayer, as a non-toxic intracellular cryopreservative for mammalian cells. Introduction of between 8 ± 3mM and 266 ± 22 mM trehalose into human erythrocytes using the membrane permeabilizing polymer PP-50 allowed investigation of the relationship between intracellular trehalose concentration, pre-freeze cell volume, and cryosurvival. Cellular cryosurvival increased approximately linearly with pre-freeze cell volume up to the normal volume of fresh cells; diminished cell survival correlated with subnormal pre-freeze cell volume in some cases even at >100mM intracellular trehalose concentration. Uptake of >200 mM trehalose in cells with near-normal cell volume facilitated enhancement of cellular cryosurvival by up to 15 ± 5%.
authors.affiliation: Department of Chemical Engineering and Biotechnology, University of Cambridge, Pembroke Street, United Kingdom.
authors.forename: Andrew L
authors.initials: AL
authors.lastname: Lynch
authors.forename: Nigel K H
authors.initials: NK
authors.lastname: Slater
mesh_terms: D001793:Blood Preservation / Q000379:methods*; D048429:Cell Size / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects; D015925:Cryopreservation / Q000379:methods*; D003451:Cryoprotective Agents / Q000494:pharmacology*; D004912:Erythrocytes / Q000187:drug effects* / Q000502:physiology; D006801:Humans; D014199:Trehalose / Q000032:analysis*
chemical_list: D003451:Cryoprotective Agents; D014199:Trehalose