Curcumin (CUR) is a major component of a dietary spice derived from the roots of Curcuma longa. It has strong antioxidant activities and hepatoprotective properties. Primary human hepatocytes are clinically used in transplantation or in bioartificial liver devices for the treatment of patients with liver failure. Fresh and cryopreserved hepatocytes are also used in vitro for the study of drugs in pharmacotoxicology. We aimed to assess whether CUR could improve human liver cell viability and prevent oxidative damage responsible for large cell loss during cell preparation. Our study showed beneficial effects of CUR (25 microM) on freshly isolated human hepatocytes, increasing significantly metabolic activity of viable attached cells when seeded with CUR for 24 h. However CUR added during the cell isolation process did not have any significant impact on cell isolation outcomes or on cryopreservation outcomes. Conversely, CUR added during the thawing of frozen cells had a negative effect on the cell attachment capacity of hepatocytes that were cryopreserved in the presence or absence of CUR. In conclusion, although having positive effects on viability and challenge of oxidative stress on cultured human hepatocytes, CUR had no beneficial effect on cell isolation or cryopreservation outcomes.