Studies on gluconeogenesis and stimulation of glycogen and protein synthesis in isolated hepatocytes in alloxan diabetic, normal fed and fasted animals.

Hepatocytes were isolated by collagenase in vitro perfusion technique. Net glucose production in isolated hepatocytes obtained from fed, fasted and alloxan diabetic rats was studied. Net glucose production from alanine, pyruvate and fructose was increased by 2-5 fold in isolated hepatocytes obtained from fasted and alloxan diabetic rats. Similar increases in the incorporation of 14C-bicarbonate into glucose was also observed. Net glucose production in isolated hepatocytes was also compared to other in vitro preparations. Net glucose production was much higher (2-5 fold) in isolated hepatocytes than that reported previously for liver slices or perfused liver. Studies on glycogen and protein synthesis show a 2 fold stimulation in the incorporation of 1-14C-glucoase into glycogen and U-14C-leucine into protein by the addition of 100 muU of insulin to isolated hepatocytes.