PMID: 2481150Dec 1, 1989Paper

Excretory function in cultured hepatocytes from griseofulvin-treated mice.

Laboratory Investigation; a Journal of Technical Methods and Pathology
H KawaharaS W French

Abstract

We determined the role of cytokeratin (CK) intermediate filaments in the excretory function of hepatocytes in cultured hepatocytes containing Mallory bodies (MBs) from the livers of griseofulvin (GF)-fed mice. Hepatocytes for primary culture were obtained from GF-fed and control mice using the 0.1% collagenase perfusion method. Each component of the cytoskeleton in cultured hepatocytes and liver frozen sections was visualized by immunofluorescence. The whole mount extraction of hepatocytes was carried out using 0.5% Triton X-100. To examine the excretory function of the bile canaliculi (BC), fluorescein diacetate and horseradish peroxidase were used as visible excretory products. Thin sections of the cultured cells were made by the "pop-off" method for electron microscopic examination. Frozen sections of livers from the GF-fed mice showed that the MBs were stained with a rat monoclonal antibody to mouse CK, but the CK filaments in the cells containing MBs did not stain. The intercellular BC were reduced in number in the livers of the GF-fed mice compared with the controls. At 3 hours after seeding, hepatocytes with MBs were not stained, but by 24 hours the CK filament network stained normally in cells containing MBs. The loss o...Continue Reading

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