Introduction: In organotypic tissue culture models, human primary bronchial epithelial cells form a pseudostratified epithelium, similar to the airway structure in vivo. Such models are cultured at airliquid interface, thus suitable for toxicity assessment of various aerosols. The complexity of the in vivo cellular response may be better captured in co-culture models, in which additional cell types, e.g. fibroblasts, were included. However, the presence of another cell type hinders the interpretation of the exposure impact on the epithelial cells. We compared the impact of a 28-minute whole combustible cigarette smoke (CS) exposure on two different organotypic bronchial models: a co-culture of the epithelial cells with fibroblasts and a mono-culture without fibroblasts. Secreted cytokines levels in the basolateral media of both models were measured 48 hours post-exposure. Additionally, transcriptomes were generated at different post-exposure time points from the epithelial cells of the mono-culture model and from the epithelial cells scraped from the co-culture model. The transcriptomics data were further analyzed using a network-based systems biology approach.