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Lung cancer is the malignancy with the highest mortality worldwide and possesses prominent histological heterogeneity. It mainly consists of three major subtypes: lung adenocarcinoma, lung squamous cell carcinoma and small cell lung cancer, along with rare pathological types such as adenosquamous carcinoma and large cell neuroendocrine carcinoma. Conventional tumor cell line models feature easy operation, rapid culture and suitability for high-throughput screening, which serve as classic in vitro tools for investigating the mechanisms underlying tumor targeted therapy. Nevertheless, genomic studies have verified that lung cancer exhibits remarkable inter-tumoral and intra-tumoral heterogeneity; phenotypic and genetic characteristics vary drastically among patients and even within distinct regions of a single tumor. Tumor cell lines alone cannot recapitulate the three-dimensional architecture and heterogeneity of primary tumors, failing to faithfully mimic the complex biological behaviors of lung cancer, which results in prominent model limitations. Patient-derived xenograft (PDX) models can well preserve the tissue architecture, genetic profiles and pathological phenotypes of human tumors, and can be stably passaged for up to 14 times, overcoming the shortcomings of cell line models. However, PDX models have inherent drawbacks: the establishment success rate is merely 30%–40%, the modeling period lasts 2–10 months, accompanied by high experimental costs and low screening throughput. These factors greatly restrict their wide application in high-throughput drug screening and large-sample mechanistic research [21]. In contrast, lung cancer organoids have a short modeling period and can be stably passaged long-term. They are capable of highly recapitulating the morphological structure, pathological features and molecular genetic phenotypes of primary lung tumors, acting as a more ideal novel in vitro tumor model. Human lung cancer organoids derived from patients’ tumor cells have been widely adopted in various cancer research fields, with core applications summarized as follows: Disease model construction: Precisely recapitulate tumor heterogeneity and microenvironmental features of lung cancer, providing a reliable in vitro system to dissect molecular mechanisms governing lung cancer initiation, malignant progression, invasion and metastasis. Drug screening and personalized therapy: Enable in vitro evaluation of the efficacy, sensitivity and toxicity of various anti-tumor drugs, offering experimental evidence and guidance for personalized precision treatment of lung cancer patients. Gene function research: Combine gene-editing technology to achieve specific knockout and knock-in of target genes, so as to accurately explore the regulatory roles of key genes in the malignant progression of lung cancer. Tumor microenvironment research: Co-culture with stromal cells including immune cells and fibroblasts to biomimetically reconstruct the tumor microenvironment, and further elucidate the interaction mechanisms between tumor cells and their microenvironment. Basic biological research: Utilize the organoid biomimetic system to investigate the development and physiological functions of normal alveolar epithelial cells, and advance the systematic understanding of lung cancer pathogenesis. Research on invasion and metastasis mechanisms: Effectively simulate the dynamic invasion and metastasis processes of lung cancer cells, providing an innovative research platform to uncover core molecular regulatory mechanisms driving malignant tumor evolution.