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.
Keywords: Cryoprotection; Electrosterically stabilized cellulose nanocrystals; Ice recrystallization inhibition.
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