The isolated hepatocyte preparation: 30 years on

A method for the preparation of intact rat hepatocytes in high yield was fi rst described in 1969. The procedure involved digestion of hepatic tissue b y perfusion of the liver with crude collagenase; later, purified collagenas e without other enzymic additions was shown to be ineffective. Recently it has been discovered that the combination of purified collagenase plus elast ase is superior to crude collagenase in that it consistently provides high yields of undamaged hepatocytes. The isolated hepatocyte preparation has pr oved particularly useful for the study of mechanisms responsible for long-r ange interactions within the cell. These can be studied over prolonged time courses and in the presence of graded concentrations of specific inhibitor s. Studies of this kind have demonstrated a close relationship between cyto plasmic metabolic hows and mitochondrial forces and have also revealed that the cytoplasmic and mitochondrial free NAD-linked redox potentials are mai ntained by energy-dependent reactions.