Successful cryopreservation of hepatocytes, especially human hepatocytes, is one of the major reasons for the recent routine application of this experimental system in drug development.   Cryopreserved human hepatocytes retain viability and metabolic capacity and are used extensively as suspension cultures to evaluate the metabolic fate (metabolic stability and metabolite profiling) of new chemical entities (NCE) during drug development. Pooled cryopreserved human hepatocytes, i.e., hepatocytes cryopreserved from several individual donors that have been thawed, pooled, and re-cryopreserved, represent the most commonly used system for routine metabolism studies. One major issue with cryopreservation, namely, the loss of the ability of the cells to be cultured, has been overcome. Now hepatocytes from both animals and humans can be cryo-preserved to retain their ability to form monolayer cultures (known as “plateable” cryopreserved hepatocytes). The use of “plateable” cryopreserved hepatocytes enhances experimental efficiency by providing an immediate supply of easily stored cells and eliminating the centrifugation steps to remove test articles after treatment (e.g. uptake and time-dependent inhibition studies), which is required for suspension cultures. Plating extends their use in applications that involve culturing   for a prolonged period (multiple days), such as evaluating metabolic stability of slowly-metabolized compounds, P450 induction, efflux transport, and hepatotoxicity. A significant advancement in the application of plateable cryopreserved hepatocytes is the evaluation of the role of metabolism-based drug toxicity on extrahepatic organs in a single test system, especially the novel Integrated Discrete Multiple Organ Co-culture (IdMOC™) system.