Isolation and morphologic characterization of bile duct epithelial cells from normal rat liver

doi:10.1016/0016-5085(89)91695-8

Gastroenterology

Volume 97, Issue 5, November 1989, Pages 1236-1247

https://doi.org/10.1016/0016-5085(89)91695-8 Get rights and content

Abstract

To study directly the functions of the cells that line the bile ducts inside the liver, we developed a new technique for isolating intrahepatic bile duct epithelial cells (IBDECs) from normal rat liver. Parenchymal and nonparenchymal cells were separated from whole liver by enzymatic digestion and mechanical disruption; subpopulations of individual nonparenchymal cells then were isolated by serial counterflow elutriation, isopycnic centrifugation, and immunoaffinity separation with a specific monoclonal antibody against an antigen on the plasma membrane of IBDECs. Using this approach, we isolated 1.2 ± 0.2 × 10⁶ (mean ± SE) viable (>95% trypan blue exclusion) cells, >95% of which were identified as IBDECs by morphologic appearance and specific cytochemical markers. The IBDECs averaged 7.4 ± 0.16 μm in diameter and retained their in situ appearance, including morphologic polarity. They appeared as single cells or as cell doublets attached by tight junctions that excluded ruthenium red. Microvilli were abundant and were restricted to the apical (i.e., luminal) domain of the plasma membrane. Coated pits were observed on both apical and basolateral cell surfaces. Internally, IBDECs contained a well-developed system of organelles, including mitochondria, Golgi, and discrete types of vesicles, such as coated vesicles, multivesicular bodies, and lysosomes. These results indicate that a highly purified suspension of viable, morphologically intact, and polar IBDECs can be prepared from normal rat liver using a novel approach that separates liver cells on the basis of size, density, and specific membrane components. The availability of such a model will allow experimental studies to be performed directly on IBDECs, an approach that has not previously been possible.

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: This work was supported by grants DK 24031 and DK 34988 from the National Institutes of Health and by the Mayo Foundation. Doctor Ishii was a recipient of a Boltz Research Fellowship Award from the Division of Gastroenterology, Mayo Medical School.

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Isolation and morphologic characterization of bile duct epithelial cells from normal rat liver

Abstract

Cell Biol Int Rep

Cell

Beitr Pathol

Exp Cell Res

Exp Cell Res

Gastroenterology

Anatomy of the liver

The intrahepatic biliary epithelium: an area of growing interest in hepatology

Semin Liver Dis

Endocytic uptake, transport, and catabolism of proteins by epithelial cells

Am J Physiol

Secretory function of the rabbit common bile duct

Am J Physiol

The contribution of the extrahepatic bile ducts to bile formation

Can J Physiol Pharmacol

Biliary physiologyin rats with bile ductular cell hyperplasia. Evidence for a secretory function of proliferated bile ductules

J Clin Invest

Two sites of bile formation as determined by mannitol and erythritol clearance in the guinea pig

J clin Invest

Permeability characteristics of bile duct in the rat

Am J Physiol

Current problems in diagnosis of biliary disease and cholestasis

Semin Liver Dis

Isolation of oval cells by centrifugal elutriation and comparison with other cell types purified from normal and preneoplastic livers

Cancer Res

Isolation and culture of biliary epithelial cells from the biliary tract fraction of normal rats

Liver

Cell types in rat liver cultures: their identification and isolation

Mol Cell Biochem

Coated pits, coated vesicles and receptor-mediated endocytosis

Nature