Preparation and characterization of isolated parenchymal cells from guinea pig liver

Abstract

A method is described for the preparation of isolated cells from guinea pig liver. This involves perfusionin situ, in the non-physiological direction, with collagenase. The cell yield was 20-30%, comparable with those from the livers of other species. The ratio of lactate dehydrogenase to glutamate dehydrogenase in the cells was similar to thatin vivo, indicating that there was negligible leakage of cytoplasmic enzymes. The concentrations of K⁺ and adenine nucleotides were initially lower than in the perfused liver; normal values were obtained on incubation, particularly in the presence of substrate. The L-lactate : pyruvate ratio is 16 : 1, close to established values. The totalβ-hydroxybutyrate : acetoacetate ratio indicates that the mitochondrial redox state is more oxidised than in the perfused liver, but the intracellular ratio is similar to that of the intact liver. Rates of gluconeogenesis and ureogenesis, are within the physiological range. Maximal gluconeogenesis from L-lactate was preceded by a lag period. L-lysine stimulated glucose production from L-lactate but did not abolish the lag phase. The effects of aminooxyacetate and octanoate on L-lactate gluconeogenesis were similar to those in the perfused liver.