Background <p>Lung cancer exhibits a high degree of heterogeneity, rendering its treatment one of the most difficult challenges. Current laboratory lung cancer models cannot fully preserve the diversity of the disease and predict drug responses. Owing to the rarity of human-derived samples, transgenic mouse models can be extensively used in fundamental research.</p> Results <p>We established a mouse lung cancer organoid (LCO) model derived from a genetically engineered mouse model that can spontaneously develop lung cancer. Morphological and molecular assays demonstrated that the lung cancer organoids retained the histological architecture and stem-like characteristics of their parental tumors. Successfully constructed lung cancer organoids were amenable for high-throughput drug screening in vitro.</p> Conclusions <p>This method can be used for the construction and identification of mouse-derived lung cancer organoids, which can be used to further comprehend the pathophysiology of lung cancer and evaluate drug responses in personalized medicine.</p>