Differentiation of Immortalized Human Airway Epithelial Cells

hTert-immortalized human normal airway tracheobronchial epithelial cells, BCi.NS1.1 (55 (link)) were maintained in ExPlus growth media (StemCell). To generate HAEC, BCi.NS1.1 were plated (7.5E4 cells/well) on rat-tail collagen type 1-coated permeable transwell membrane supports (6.5mm; Corning Inc), immersed in ExPlus growth media in both the apical and basal chamber. Upon reaching confluency, media in the apical chamber was removed (airlift), and media in the basal chamber was changed to Pneumacult ALI maintenance media (StemCell). Pneumacult ALI maintenance media was changed every two days for approximately 6 weeks to form differentiated, polarized cultures that resemble in vivo pseudostratified mucociliary epithelium.

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Lokugamage K.G., Hage A., de Vries M., Valero-Jimenez A.M., Schindewolf C., Dittmann M., Rajsbaum R, & Menachery V.D. (2020). Type I interferon susceptibility distinguishes SARS-CoV-2 from SARS-CoV.. bioRxiv.

Publication Preprint 2020

ExPlus growth media rat-tail collagen type 1-coated permeable transwell membrane supports Pneumacult ALI maintenance media

Cells Collagen type 1 Epithelial cells Epithelium Growth media Human Membrane Permeable Stemcell Tail

Corresponding Organization : The University of Texas Medical Branch at Galveston

Other organizations : New York University

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Variable analysis

independent variables

Plating of BCi.NS1.1 cells (7.5E4 cells/well) on rat-tail collagen type 1-coated permeable transwell membrane supports
Removal of media in the apical chamber (airlift)
Change of media in the basal chamber to Pneumacult ALI maintenance media

dependent variables

Formation of differentiated, polarized cultures that resemble in vivo pseudostratified mucociliary epithelium

control variables

BCi.NS1.1 cell line
ExPlus growth media in both the apical and basal chamber until confluency
Pneumacult ALI maintenance media changed every two days for approximately 6 weeks

controls

Positive control: Not explicitly mentioned
Negative control: Not explicitly mentioned

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