Culture and characterization of rat middle-ear epithelium

Acta Otolaryngol. May-Jun 1986;101(5-6):453-66. doi: 10.3109/00016488609108632.

Abstract

This study was performed to design a method for the culture of rat middle-ear epithelium and to apply the method to investigate the characteristics of this epithelium. Culture of explants of middle-ear epithelium in the presence of the epidermal growth factor was successful, whereas serial cultivation required 3T3 feeder cells in addition to the epidermal growth factor. Cultured middle-ear epithelium was studied by phase-contrast microscopy, transmission and scanning electron microscopy, and combined light and scanning electron microscopy (LM/SEM). These techniques showed similarity between the cultured and the natural middle-ear epithelium. Explants and outgrowths showed both flat polygonal and ciliated epithelial cells. In serial cultivation, however, only the first of these cell types was observed. Frequently, a single primary cilium was found on the cell surface. Transmission electron microscopy showed cross-linked envelopes whose formation was promoted by ionophore X537A. Cytokeratin was demonstrated by immunoblotting, immunofluorescence, and immunoperoxidase methods, using an anti-cytokeratin monoclonal antibody. The model described here permits study of the differentiation of middle-ear epithelium in vitro and may be of future value for the study of chronic middle-ear diseases.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cell Membrane / ultrastructure
  • Cells, Cultured
  • Ear, Middle / ultrastructure*
  • Epithelial Cells
  • Epithelium / ultrastructure
  • Keratins / analysis
  • Male
  • Microscopy, Electron
  • Microscopy, Electron, Scanning
  • Microscopy, Phase-Contrast
  • Microvilli / ultrastructure
  • Rats
  • Rats, Inbred Strains
  • Staining and Labeling

Substances

  • Keratins