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Updated: Aug 7, 2026
Generating Primary Fibroblast Cultures from Mouse Ear and Tail Tissues
Published on: January 10, 2016
Identification of specific gene expression profiles in fibroblasts derived from middle ear cholesteatoma
Mamoru Yoshikawa1, Hiromi Kojima, Kota Wada
1Department of Otorhinolaryngology, Jikei University School of Medicine, Tokyo, Japan. yoshikawa@jikei.ac.jp
Objective:
To investigate the role of fibroblasts in the pathogenesis of cholesteatoma.
Design:
Tissue specimens were obtained from our patients. Middle ear cholesteatoma-derived fibroblasts (MECFs) and postauricular skin-derived fibroblasts (SFs) as controls were then cultured for a few weeks. These fibroblasts were stimulated with interleukin (IL) 1alpha and/or IL-1beta before gene expression assays. We used the human genome U133A probe array (GeneChip) and real-time polymerase chain reaction to examine and compare the gene expression profiles of the MECFs and SFs.
Subjects:
Six patients who had undergone tympanoplasty.
Results:
The IL-1alpha-regulated genes were classified into 4 distinct clusters on the basis of profiles differentially regulated by SF and MECF using a hierarchical clustering analysis. The messenger RNA expressions of LARC (liver and activation-regulated chemokine), GMCSF (granulocyte-macrophage colony-stimulating factor), epiregulin, ICAM1 (intercellular adhesion molecule 1), and TGFA (transforming growth factor alpha) were more strongly up-regulated by IL-1alpha and/or IL-1beta in MECF than in SF, suggesting that these fibroblasts derived from different tissues retained their typical gene expression profiles.
Conclusions:
Fibroblasts may play a role in hyperkeratosis of middle ear cholesteatoma by releasing molecules involved in inflammation and epidermal growth. These fibroblasts may retain tissue-specific characteristics presumably controlled by epigenetic mechanisms.
Insights
Middle ear cholesteatoma fibroblasts (MECFs) show distinct gene expression compared to skin fibroblasts (SFs). MECFs may contribute to cholesteatoma pathogenesis through inflammation and epidermal growth signaling.
Area of Science:
- Otolaryngology
- Dermatology
- Molecular Biology
Background:
- Cholesteatoma is a skin-related condition of the middle ear.
- Fibroblasts are key cells in tissue repair and inflammation.
Purpose of the Study:
- To investigate the role of fibroblasts in cholesteatoma pathogenesis.
- To compare gene expression profiles of cholesteatoma-derived fibroblasts (MECFs) and skin-derived fibroblasts (SFs).
Main Methods:
- Fibroblast cultures from middle ear cholesteatoma (MECFs) and postauricular skin (SFs).
- Stimulation with interleukin (IL)-1alpha and/or IL-1beta.
- Gene expression analysis using human genome U133A probe array and real-time PCR.
Main Results:
- Hierarchical clustering identified distinct gene expression profiles.
- MECFs showed stronger upregulation of LARC, GMCSF, epiregulin, ICAM1, and TGFA compared to SFs upon IL-1 stimulation.
- These findings suggest tissue-specific gene expression patterns are retained.
Conclusions:
- Fibroblasts may contribute to cholesteatoma hyperkeratosis via inflammatory and growth-related molecules.
- Tissue-specific fibroblast characteristics are likely maintained through epigenetic mechanisms.
