Abstract
The upper respiratory tract is one of the first lines of defence against inhaled pollutants or toxic material. Two of the essential elements of this protection are coordinated ciliary movement and barrier function of the bronchial epithelial cells. Normal human bronchial epithelial cells (NHBE) from primary culture can be co‐cultured with human lung fibroblasts (Wi‐38) as a bilayer on Transwell filter plates. Using the air‐liquid interface (ALI) technique we show that NHBE form a pseudostratified epithelium and express both tight (TJ) and adherens junction (AJ) proteins after 14 to 17 days in vitro. Mucus production and cilia formation developed within 21 days. Additionally, these cilia showed a beat frequency about 16 to 20 Hz. The differentiated state of the bronchial cells was maintained up to three months, correlating with average transepithelial resistance‐values between 600‐800 ohm.cm2 and prolonged ciliary beating. Exposure to TNF‐alpha and IFN‐gamma induced significant reduction in barrier functions (reduced expression of ZO‐1, occludin, E‐cadherin, beta‐catenin : immunofluorescence, Western blot, RT‐PCR; increased permeability for FITC dextrans) and ciliary beat frequency. These data demonstrate a relevant co‐culture model to investigate pathomechanisms of proximal respiratory tract injury. (Supported by the German Defence Ministry)
Published Version
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