Characterization of a temperature-sensitive mouse middle ear epithelial cell line

Katsuyuki Tsuchiya, Youngki Kim, Frank G Ondrey, Jizhen Lin

Research output: Contribution to journalArticlepeer-review

23 Scopus citations


Conclusion. Temperature-sensitive mouse middle ear epithelial cells have been successfully established and characterized. Objective. Temperature- sensitive middle ear epithelial cell lines are essential for pathophysiologic studies of otitis media. They are useful for studying the pathogen-host interaction, receptor identification, signal transduction, cytokine/mucin production and cellular responses, especially for cell proliferation and differentiation. The purpose of this study was to establish a large T-antigen [simian virus 40 A-gene (SV40)] mutant-immortalized mouse middle ear epithelial cell line for otitis media studies. Material and methods. Primary culture of middle ear epithelial cells was established from the middle ear mucosa of an Immortomouse. The cells were transduced by a temperature-sensitive large T-antigen mutant and cultured for >50 passages. The expression of mRNA transcripts and proteins for epithelial cells was characterized by reverse transcriptase polymerase chain reaction (RT-PCR) and immunohistochemistry. The temperature-sensitive properties of cells cultured at 33°C and 39°C were evaluated using 3H-thymidine incorporation, Trypan blue exclusion and peroxisome proliferator-activated receptor-γ activity. Results. Immortalized middle ear epithelial cells demonstrated a cobblestone-like monolayer culture. The cells expressed mucosal cell markers such as mucins, keratins and collagens. They proliferated at 33°C when the SV40 antigen was active and differentiated at 39°C when the SV40 antigen was inactive.

Original languageEnglish (US)
Pages (from-to)823-829
Number of pages7
JournalActa Oto-Laryngologica
Issue number8
StatePublished - 2005

Bibliographical note

Funding Information:
We thank Dr Bing Zhou for providing us with the SV40 vector, Dr Masahiro Komori for reading the manuscript and Eileen P. Schlentz for editorial assistance in the preparation of the manuscript. This work was supported in part by grant Nos. R01DC03433 and P30DC04660 from the National Institutes of Health, grant No. R21 DC005846 from the National Institute of Deafness and Other Communication Disorders and by the Lions 5M Multiple District Hearing Foundation.


  • Immortalization
  • Luciferase assay
  • Middle ear epithelial cells
  • Mouse
  • Reverse transcriptase polymerase chain reaction
  • Tissue culture


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