Chronic lung diseases such as chronic obstructive pulmonary disease, cystic fibrosis and idiopathic lung disease are incurable. Epithelial senescence, a state of dysfunctional cell cycle arrest, contributes to the progression of such diseases. We have used lung organoids (bronchospheres) to test active substances and polyinosinic:polycytidylic acid (pIC) for senescence, toxicity and inflammation, with pIC mimicking viral infection. We treated basal cells for five consecutive days with pIC, the senolytic drug quercetin, the combination of pIC and quercetin, and with control medium. The basal cells were washed and cultured in culture media for additional 2 to 7 days. Basal cells treated with pIC showed a senescent phenotype and had a reduced ability to form motile organoids. Pharmaceutical compounds such as quercetin and dasatinib antagonized doxorubicin-induced senescence and selectively decreased the release of epithelial mediators without compromising organoid integrity. Our results indicate that exogenous or viral stressors induce a persistent senescent phenotype in respiratory epithelial cells, which is associated with a reduced capacity for cell differentiation.