Ciliated differentiation of rabbit tracheal epithelial cells in vitro

Primary cultures of rabbit tracheal epithelial (RbTE) cells have been perfo rmed in two different ways. Quantitative analysis of both proliferative cap acities and ciliated differentiation process were carried out using epithel ial cell cultures from tracheal explants and from dissociated tracheal epit helial cells in air-liquid interface conditions. We show that both alpha- a nd beta-tubulins from RbTE cells are polyglutamylated and that this posttra nslational modification is restricted to cilia axonemes and centrioles of n on-ciliated cells. A monoclonal antibody raised against polyglutamylated tu bulins was used to quantify the proportion of ciliated cells. Even though e pithelial cells from outgrowths obtained by the explant technique highly pr oliferated during the first days of culture, no ciliated differentiation oc curred. On the other hand, using air-liquid interface conditions after prol iferation of dissociated cells, we could observe and quantify a ciliated ce ll differentiation in vitro by both Western blot and flow cytometric analys is. The specific detection and quantification of ciliated cells open the wa y for the biochemical and molecular characterization of centriolar componen ts during ciliated differentiation.