Differentiation of Airway Epithelial Cells

hTert-immortalized human normal airway tracheobronchial epithelial cells, BCi.NS1.1 (58 (link)), were maintained in ExPlus growth media (StemCell). To generate HAEC, BCi.NS1.1 cells were plated (7.5E4 cells/well) on rat-tail collagen type 1-coated permeable transwell membrane supports (6.5 mm; Corning Inc.), immersed in ExPlus growth media in both the apical and the basal chambers. Upon reaching confluence, the medium in the apical chamber was removed (airlift), and the medium in the basal chamber was changed to Pneumacult ALI maintenance media (StemCell). Pneumacult ALI maintenance medium was changed every 2 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. Journal of Virology, 94(23), e01410-20.

Publication 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 Human Membrane Permeable Stemcell Tail

Corresponding Organization :

Other organizations : The University of Texas Medical Branch at Galveston, New York University

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Variable analysis

independent variables

Culture conditions: Cells were initially cultured in ExPlus growth media, then switched to Pneumacult ALI maintenance media after reaching confluence and removing the medium from the apical chamber (airlift)

dependent variables

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

control variables

Cell line: hTert-immortalized human normal airway tracheobronchial epithelial cells, BCi.NS1.1
Seeding density: 7.5E4 cells/well
Extracellular matrix: Cells were plated on rat-tail collagen type 1-coated permeable transwell membrane supports
Membrane size: 6.5 mm

controls

Positive control: None specified
Negative control: None specified

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