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Characterization and response to interleukin 1 and tumor necrosis factor of immortalized murine biliary epithelial
1Department of Pediatrics, Sainte Justine Hospital Research Center, University of Montreal, Quebec, Canada.
Insights
Researchers established pure, immortalized murine intrahepatic biliary epithelial cells. This breakthrough enables detailed studies of these cells in liver diseases.
Area of Science:
- Hepatology
- Cell Biology
- Immunology
Background:
- Biliary epithelial cells are crucial in immune-mediated liver diseases.
- Their exact role is unclear due to challenges in isolating pure cell populations.
Purpose of the Study:
- To establish pure, immortalized clones of murine intrahepatic biliary epithelial cells.
- To facilitate research into the function and behavior of these cells.
Main Methods:
- Utilized transgenic mice with a thermosensitive SV40 gene (TsA58) under MHC class I promoter control.
- Employed countercurrent centrifugal elutriation and clonal dilution for cell isolation.
- Established conditionally immortalized intrahepatic bile duct cells.
Main Results:
- Obtained immortalized cell clones expressing cytokeratin 19, forming duct-like structures.
- Electron microscopy confirmed well-differentiated and polarized cells.
- Cells responded to epidermal growth factor, IL-1α, and TNFα, expressing receptors for these cytokines.
Conclusions:
- Availability of unlimited pure bile duct cells is now possible.
- These cells mimic normal biliary epithelial cells, enabling rigorous functional studies.
- Facilitates advanced research into biliary epithelial cell behavior in liver disease pathogenesis.
Background & Aims:
Biliary epithelial cells are the target of numerous immune-mediated liver diseases, yet their role in pathogenesis remains unclear because of difficulties in obtaining pure preparations. The aim of this study was to establish pure clones of immortalized murine intrahepatic biliary epithelial cells.
Methods:
The transgenic mouse harboring the SV40 thermosensitive immortalizing mutant gene TsA58 under the control of the major histocompatibility complex class I promoter was used to establish conditionally immortalized intrahepatic bile duct cells by countercurrent centrifugal elutriation and clonal dilution.
Results:
Immortalized clones of cells expressing cytokeratin 19, which organized themselves into ductlike structures, were obtained. On electron-microscopic sections, cells were well differentiated and polarized. Cells proliferate in response to epidermal growth factor, interleukin 1 alpha, and tumor necrosis factor alpha. Using the reverse-transcriptase polymerase chain reaction technique, these cells were found to contain messenger RNA, which encodes for the interleukin 1 and tumor necrosis factor receptors.
Conclusions:
The availability of unlimited numbers of pure bile duct cells that behave in an identical fashion to biliary epithelial cells from "normal" mice will allow for more rigorous studies of the behavior and function of this epithelium.
