The quality of the hepatocytes used for clinical cell transplantation is very important, and depends to a large extent on the nature of the tissue used for isolation. The collagenase perfusion technique to isolate hepatocytes from animal livers has been further developed for isolation of human hepatocytes. As the donor organ pool is a scarce resource, marginal livers unsuitable for transplantation and segments from reduced grafts remain the main source of tissue for cell isolation. Use of livers from non-heart beating donors and foetal livers may further increase the tissue pool. With the limited supply of available tissue, improvements in the cryopreservation protocols are required to maintain cell viability on thawing and establish hepatocyte banks.