Enhanced Metabolic Function of Human Hepatocytes Cryopreserved with Low Concentration Me₂SO and Polyol Additives at –80°C

The metabolic function of cryopreserved cells, in addition to cell viability after thawing, is an important parameter in any successful cryopreservation protocol. Dimethyl sulfoxide (Me₂SO) is known to affect the differentiation of recovered cells. In this study, we report that sugars and sugar alcohols increases cell recovery, and also improves the metabolic function of human hepatocytes that are cryopreserved using low concentration Me₂SO (5%). Three sugars (glucose, sucrose, and trehalose) and three sugar alcohols (xylitol, maltol, and sorbitol) have been tested. Cell viability after thaw and 24-h post-thaw attachment rate of cryopreserved human hepatocytes were assessed. Post-thaw metabolic activities (albumin, glucose, urea content) were measured, and cell proliferation was observed with inverted microscope. Cell viability, post-thaw attachment rate and metabolic activity of cryopreserved hepatocytes are enhanced by the addition of 0.4M sorbitol into 5% Me₂SO solution. The study concludes that 5% Me₂SO + 0.4M sorbitol can replace the 10% Me₂SO method for cryopreservation of human hepatocytes at -80°C freezer. The new solution may reduce the side effects on the patients and improve the safety of using cryopreserved hepatocytes.