Extracellular Antifreeze Protein Significantly Enhances the Cryopreservation of Cell Monolayers

, from 25% to over 60%, whereas intracellularly delivered AFPIII showed less benefit. Interestingly, the antifreeze protein was less effective when used in suspension cryopreservation of the same cells, suggesting that the cryopreservation format is also crucial. These observations show that, in the discovery of macromolecular cryoprotectants, intracellular delivery of ice recrystallization inhibitors may not be a significant requirement under "slow freezing" conditions, which will help guide the design of new biomaterials, in particular, for cell storage.