Biopolymer mediated trehalose uptake for enhanced erythrocyte cryosurvival.

Abstract

A biopolymer has been shown to facilitate efficient delivery of trehalose, a bioprotectant normally impermeable to the phospholipid bilayer, into ovine erythrocytes. Cellular uptake of trehalose was found to be dependent on polymer pendant amino acid type and degree of grafting, polymer concentration, pH, external trehalose concentration, incubation temperature and time. Optimization of these parameters yielded an intracellular trehalose concentration of 123 +/- 16 mM and concomitant improvement of erythrocyte cryosurvival of up to 20.4 +/- 5.6%. Intracellular trehalose was shown to impart cellular osmoprotection up to an external osmolarity of 230 mOsm and increased osmotic sensitivity above this threshold. Biopolymer mediated membrane permeability was shown to be rapidly and completely reversible via washing with phosphate buffered saline.