Hepatocyte transplantation for acute liver failure and metabolic liver disorders represent a potential alternative to traditional orthotopic liver transplantation (OLT). Additionally, cultured primary hepatocytes are an important tool for in vitro pharmacological studies. Advances in clinical hepatocyte transplantation and drug studies depend on the development of standardized isolation and culturing protocols as well as high-quality enzymes for liver tissue digestion. The isolation of primary hepatocytes for clinical applications must comply with current Good Manufacturing Practices (cGMP) regulations which may include the use of digestion enzymes produced to cGMP standards.