Barrier functions and paracellular integrity in human cell culture models of the proximal respiratory unit.
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.
Pohl
Christine
C
Institute of Pathologie, Johannes Gutenberg University Mainz, Germany. pohlc@uni-mainz.de
Hermanns
M Iris
MI
Uboldi
Chiara
C
Bock
Michaela
M
Fuchs
Sabine
S
Dei-Anang
Jasmin
J
Mayer
Eckhard
E
Kehe
Kai
K
Kummer
Wolfgang
W
Kirkpatrick
Charles James
CJ