Comparative analysis of proteins labelled with [35S]methionine in the liver in vivo and in freshly isolated and short-term-cultured hepatocytes in vitro

[³⁵S]Methionine-labelled liver proteins, analysed by one- or two-dimensional gel electrophoresis showed a strikingly similar pattern whether synthesized in vivo or by freshly isolated hepatocytes. In contrast, major qualitative and quantitative differences were observed with the patterns of labelled proteins found in cultured hepatocytes. The changes detectable very early (within 1 h) in culture affected preferentially the synthesis of cytoskeleton proteins (cytokeratins, actin, myosin), which was dramatically increased. Physical factors like cell attachment appear to be responsible for these changes which, however, occurred more rapidly in the presence of serum. Freshly isolated hepatocytes and short-term-cultured cells responded similarly to insulin and glucagon, which respectively increased and decreased the labelling of the whole set of cellular and exported proteins. Glucocorticoids caused either an increase or a decrease in the labelling of several proteins, but the effects were detectable only under chronic exposure of cultured hepatocytes. Based on these results, freshly isolated hepatocytes appear more representative of the liver in vivo than cultured hepatocytes, and therefore seem more suitable for short-term studies. However, cultured hepatocytes can be used for long-term studies since they maintain many specific liver functions and remain hormonally sensitive.