摘要
Cryopreservation plays an essential role in biobanking and cell therapy,but the physiological toxicity of traditional cryoprotectants such as glycerol and dimethyl sulfoxide(DMSO)has raised safety issues for biomedical applications.Trehalose,a nonreducing disaccharide that accumulates in desiccation-or cold-tolerant organisms,has been considered as a biocompatible cryoprotectant.Herein,a naturally occurring membrane-active cationic peptide,melittin,was utilized to facilitate membraneimpermeable trehalose entry into cells for effective cell cryopreservation.Poly(ethylene glycol)and poly(vinyl pyrrolidone)were applied as macromolecular protectants to improve the stabilization of cell membranes.Upon the optimal protocol,the postthaw recovery of human red blood cells in freezing bags at a hematocrit of~50%could achieve 82.9%with favorable cell morphologies and physiological functions.Furthermore,the cryosurvival of L929 fibroblasts reached 84.3%compared to the conventional method using 10%(v/v)DMSO.In short,this work by using trehalose and melittin provides a biocompatible solvent-free approach for long-term cryostorage of cells.
