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Primary basal cell expansion Primary human basal airway epithelial cells from tracheal digests were expanded at 37 C on NIH 3T3 fibroblast feeders in F-media (67.5% DMEM-F, 25% Hams F-12, 7.5% FBS, 1.5 mM l-glutamine, 25 ng/mL hydrocortisone, 12 5 ng/mL EGF, 8.6 ng/mL cholera toxin, 24 g/mL Adenine, 0.1% insulin, 75 U/mL pen/strep) with ROCK1 Inhibitor (RI, 16 g/mL), and antibiotics (1.25 g/mL amphotericin B, 2 g/mL fluconazole, 50 g/mL gentamicin) 67 , and cryopreserved in freeze media upon initial passaging (P1)

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