title: Barrier functions and paracellular integrity in human cell culture models of the proximal respiratory unit.
abstract: Airway epithelial cells provide a barrier to the translocation of inhaled materials. Tight (TJ) and adherens junctions (AJ) play a key role in maintaining barrier functions, and are responsible for the selective transport of various substances through the paracellular pathway. In this study we compared a bronchial cell line (16HBE14o-) and primary bronchial cells (HBEC), both cocultivated with the fibroblast cell line Wi-38, with respect to their structural differentiation and their reaction to cytokine stimulation. HBEC formed a pseudostratified epithelial layer and expressed TJ and AJ proteins after 2 weeks in coculture. Mucus-producing and ciliated cells were found within 24 days. Additionally, a beating activity of the ciliated HBEC (14-19Hz) could be detected. 16HBE14o- in coculture showed a multilayered growth without differentiation to a pseudostratified airway epithelium. Simultaneous exposure to TNF-alpha- and IFN-gamma-induced significant changes in barrier function and paracellular permeability in the cocultures of HBEC/Wi-38 but not in the 16HBE14o-/Wi-38. In summary, HBEC in coculture mimic the structure of native polarized bronchial epithelium showing basal, mucus-producing and ciliated cells. Our system provides an opportunity to examine the factors that influence barrier and mucociliary function of bronchial epithelium within a time frame of 3 weeks up to 3 months in an in vivo-like differentiated model.
authors.affiliation: Institute of Pathologie, Johannes Gutenberg University Mainz, Germany. pohlc@uni-mainz.de
authors.forename: Christine
authors.initials: C
authors.lastname: Pohl
authors.forename: M Iris
authors.initials: MI
authors.lastname: Hermanns
authors.forename: Chiara
authors.initials: C
authors.lastname: Uboldi
authors.forename: Michaela
authors.initials: M
authors.lastname: Bock
authors.forename: Sabine
authors.initials: S
authors.lastname: Fuchs
authors.forename: Jasmin
authors.initials: J
authors.lastname: Dei-Anang
authors.forename: Eckhard
authors.initials: E
authors.lastname: Mayer
authors.forename: Kai
authors.initials: K
authors.lastname: Kehe
authors.forename: Wolfgang
authors.initials: W
authors.lastname: Kummer
authors.forename: Charles James
authors.initials: CJ
authors.lastname: Kirkpatrick