title: Culture of rabbit middle ear epithelial cells. A method for primary culture and subculture with identification, characterization and growth specification.
abstract: During the last decade middle ear epithelium has been cultured from various species. Until now, subcultivation has been achieved only with the use of a feeder-cell layer or conditioned medium. These factors are possible confounders in the in vitro model. On the other hand, subcultivation is necessary for exact quantitative studies. We present a reproducible culture method allowing subcultivation without feeder-cells or conditioned medium. The main features in our method are a low-serum, hormone-supplemented medium, an incubation temperature of 34 degrees C, fixation of explants, gentle trypsinization and replating with high cell density. Cells were identified by immunohistochemistry through a battery of monclonal antibodies. The percentage of epithelial cells in the subculture was 99.2%. To our knowledge, this is the first report describing subcultivation of middle ear epithelial cells exclusively in a completely controlled environment. These are optimal circumstances for future investigation and quantification of various factors influencing proliferation and differentiation of middle ear epithelium.
authors: Ear-, Nose and Throat Department, Aarhus University Hospital, Denmark.
authors: L P
authors: LP
authors: Schousboe
authors: T
authors: T
authors: Ovesen
authors: P D
authors: PD
authors: Ottosen
authors: T
authors: T
authors: Ledet
authors: O
authors: O
authors: Elbrönd
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal; D018929:Cell Culture Techniques; D004432:Ear, Middle / Q000166:cytology*; D004847:Epithelial Cells; D005260:Female; D007150:Immunohistochemistry; D011817:Rabbits
chemical_list: D000911:Antibodies, Monoclonal