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Characterization of a temperature-sensitive mouse middle ear epithelial cell line
Katsuyuki Tsuchiya1, Youngki Kim, Frank G Ondrey
1Department of Otolaryngology, Otitis Media Research Center, University of Minnesota School of Medicine, Minneapolis, Minnesota, USA.
Acta Oto-Laryngologica
|September 15, 2005
Summary
Temperature-sensitive mouse middle ear epithelial cells were established for otitis media research. These cells proliferate at 33°C and differentiate at 39°C, aiding in pathogen-host interaction studies.
Area of Science:
- Otorhinolaryngology
- Cell Biology
- Molecular Biology
Background:
- Temperature-sensitive middle ear epithelial cell lines are crucial for studying otitis media.
- These cell lines facilitate research into pathogen-host interactions, receptor identification, and cellular responses.
Purpose of the Study:
- To establish a temperature-sensitive, immortalized mouse middle ear epithelial cell line using a simian virus 40 A-gene (SV40) large T-antigen mutant.
- To create a valuable tool for otitis media research, enabling detailed studies of cellular mechanisms.
Main Methods:
- Primary middle ear epithelial cells were cultured from Immortomice.
- Cells were immortalized using a temperature-sensitive SV40 large T-antigen mutant and cultured for over 50 passages.
- Cellular characteristics, temperature-sensitive properties, and marker expression were assessed using RT-PCR, immunohistochemistry, and functional assays.
Main Results:
- The established immortalized cells formed a cobblestone-like monolayer and expressed epithelial markers (mucins, keratins, collagens).
- Cells exhibited temperature-dependent behavior: proliferation at 33°C (SV40 active) and differentiation at 39°C (SV40 inactive).
Conclusions:
- Successfully established and characterized temperature-sensitive mouse middle ear epithelial cells.
- These cells provide a robust model for investigating otitis media pathogenesis and cellular responses.