Preparation of hepatocytes.

This unit describes a process for isolation of intact, viable hepatocytes for use in integrated drug metabolism studies. Isolated hepatocytes are increasingly being used as a biological system for studying the in vitro metabolism of xenobiotics. The isolation of hepatocytes from laboratory animals and nontransplantable human livers donated for research activities involves a two-step enzymatic digestion of the liver tissue. Two methods for the perfusion of liver tissue are included: in situ perfusion and isolation of hepatocytes and perfusion of excised tissue. The basic protocol also includes suggestions for designing drug metabolism experiments using hepatocytes. The final protocol describes cryopreservation and long-term storage of isolated hepatocytes.