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

Motoyasu Ishii, Benjamin Vroman, Nicholas F La Russo

Research output: Contribution to journalArticle

177 Citations (Scopus)

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.

Original languageEnglish (US)
Pages (from-to)1236-1247
Number of pages12
JournalGastroenterology
Volume97
Issue number5
StatePublished - 1989

Fingerprint

Intrahepatic Bile Ducts
Bile Ducts
Epithelial Cells
Liver
Isopycnic Centrifugation
Cell Membrane
Coated Vesicles
Multivesicular Bodies
Ruthenium Red
Trypan Blue
Tight Junctions
Microvilli
Lysosomes
Organelles
Digestion
Suspensions
Mitochondria
Monoclonal Antibodies
Antigens
Cell Line

ASJC Scopus subject areas

  • Gastroenterology

Cite this

Isolation and morphologic characterization of bile duct epithelial cells from normal rat liver. / Ishii, Motoyasu; Vroman, Benjamin; La Russo, Nicholas F.

In: Gastroenterology, Vol. 97, No. 5, 1989, p. 1236-1247.

Research output: Contribution to journalArticle

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