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Thus, a 96-well microplate system was developed where primary human small airway epithelial (hSAE) cells were cultured at an air\u2013liquid interface (ALI). After four weeks of ALI culture, a pseudostratified epithelium containing basal, club, goblet and ciliated cells was produced. The 96-well ALI cultures displayed a cellular composition, ciliary beating frequency, and intercellular tight junctions similar to 24-well conditions. A novel custom-made device for 96-parallelized transepithelial electric resistance (TEER) measurements, together with dextran permeability measurements, confirmed that the 96-well culture developed a tight barrier function during ALI differentiation. 96-well hSAE cultures were responsive to transforming growth factor \u03b21 (TGF-\u03b21) and tumor necrosis factor \u03b1 (TNF-\u03b1) in a concentration dependent manner. 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