Electrosterically stabilized cellulose nanocrystals demonstrate ice recrystallization inhibition and cryoprotection activities

Abstract Ice recrystallization inhibitors have emerged as novel cryoprotectants to improve cell viability for cryopreservation. Nanocelluloses were identified as new materials for ice recrystallization inhibition (IRI); however, conventional nanocelluloses aggregate and lose IRI activity at high ionic strengths, which limit their application as cryoprotectants. In this study, we synthesized a novel group of nanocelluloses - electrosterically stabilized cellulose nanocrystals (ECNCs), which remained dispersed and IRI-active at high ionic strengths. ECNCs improved the post-thaw viability of HCT-116 colorectal cancer cells in slow/fast freezing-slow thawing protocols in the presence of 1–20% v/v dimethyl sulfoxide (DMSO), as well as in slow/fast freezing-fast thawing protocols at reduced DMSO concentrations. The effectiveness in cryoprotection did not match the IRI activity in ECNCs, polyethylene glycol (PEG), and polyvinyl alcohol (PVA); and in ECNCs with different surface charge densities. Overall, ECNCs demonstrated IRI and cryoprotection activities, but the mechanism of cryoprotection remains unknown.