[Isolation of periportal and centrolobular hepatocytes by isopycnic centrifugation]

Suspensions of rat hepatocytes were separated by Ficoll discontinuous density gradient into four fractions. About 85% of the total quantity of cells sedimented within the range from 1.044 to 1.126 g/ml. The activity of key enzymes of glycolysis and gluconeogenesis were measured to determine the acinar origin of hepatocyte subpopulations. The activity of the gluconeogenic enzyme, phosphoenolpyruvate carboxykinase, in hepatocytes with the density of 1.044 g/ml was twice as high as in hepatocytes with the density of 1.073 g/ml. In contrast, glycolytic enzyme, hexokinase, was 3 times more active in heavy than in light cells. The results indicate that light and heavy cells correspond to periportal and centrilobular hepatocytes, respectively.