Optimisation of the cryopreservation of primary hepatocytes.

The use of cryopreserved hepatocytes has increased in the last decade due to the improvement of the freezing and thawing methods and has even achieved acceptance by the U.S. Food and Drug Administration for use in drug-metabolising enzyme induction studies. This chapter provides an overview of the theories behind the process of cryopreservation and some of the most important advances which have led to the ability to cryopreserve hepatocytes, which when thawed retain functions similar to fresh cells. Parameters such as cell density, cryoprotectants and freezing media should be considered as well as storage conditions and thawing techniques. Special emphasis is placed on human hepatocytes but information for the cryopreservation of animal hepatocytes is also described. Finally, a suggested method for optimising cryopreservation method is outlined.