Modulation of expression of intermediate filaments during the development of established rat liver cell lines

A number of clear epithelial-like cell lines were established from the liver of fetal and neonatal rats. The intermediate filaments of these cells were investigated using polyclonal prekeratin antisera, monoclonal antibodies against a cytokeratin subfamily and vimentin by means of the indirect immunofluorescence technique and SDS-PAGE analysis. Changes in the expression of cytokeratin filaments were found during the evolution of permanent cell lines. Cells positively stained for cytokeratins could be seen near to other cells which were negative. In early passages (up to the 40th) nearly all cells were strongly stained by the different keratin antibodies. During the following subcultivation the pattern of staining considerably changed. In FRL and NP-RL cell lines keratin-negative cells could already be observed in the early passages, rapidly increasing in the later passages. Compared to this, vimentin-staining of all cells remained constant in its morphological expression. The keratin filaments were seen in thick fiber bundles arranged particularly in the perinuclear ring as well as in finer networks throughout the cytoplasm. Every cell in the established lines showed their very individual staining pattern. The vimentin filaments extended to the whole cytoplasm up to the cell margin. Our observations demonstrate the variability of the system of keratin filaments in established epithelial-like liver cells under cultural conditions.