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Confluent monolayers of bile duct epithelial cells with tight junctions
Author(s) -
Okamoto Hiromasa,
Ishii Motoyasu,
Mano Yutaka,
Igarashi Takehiko,
Ueno Yoshiyuki,
Kobayashi Koju,
Toyota Takayoshi
Publication year - 1995
Publication title -
hepatology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 5.488
H-Index - 361
eISSN - 1527-3350
pISSN - 0270-9139
DOI - 10.1002/hep.1840220125
Subject(s) - tight junction , paracellular transport , in vivo , monolayer , cell junction , bile duct , in vitro , horseradish peroxidase , biliary tract , chemistry , ruthenium red , cell culture , microbiology and biotechnology , biology , biophysics , cell , biochemistry , medicine , permeability (electromagnetism) , membrane , enzyme , calcium , genetics , organic chemistry
The culture of fully differentiated intrahepatic bile duct epithelial cells (IBDECs) to use as a model for the in vivo intrahepatic biliary tract has not been established. IBDECs from normal rat livers were grown on a collagen‐coated permeable filter and formed a confluent monolayer 7 days after being plated. Positive reactions for cytokeratin‐19 and retained gamma‐glutamyl transpeptidase (GGTP) activity were shown. The transepithelial electrical resistance between the apical and the basolateral compartment culture chambers increased with the culture age and plateaued after the 7th day. The resulting cultured cells displayed a number of characteristics. (1) The cells formed a thin, continuous mono‐layer and displayed microvilli on the apical surface and junctional complexes between the cells, consistent with in vivo IBDECs. (2) Cells cultured for more than 7 days prevented the passage of horseradish peroxidase (HRP) and ruthenium red through paracellular pathways. (3) Seven‐day‐old cultures displayed a mean transepithelial electrical resistance of 137.3 Ωcm<2>, which decreased by 27.1% from its initial level after cell treatment with ethylenediaminetetraacetic acid (EDTA). These results indicate that confluent IBDEC monolayers are well differentiated and polarized with tight junctions (TJs) between the cells. These cell monolayers can provide a useful and relevant model for the in vitro study of various in vivo bile duct phenomena. (H EPATOLOGY 1995; 22:153–159.)