A427 cells originate from lung tissue, specifically a carcinoma, exhibit epithelial morphology and grow adherently. A427 cells have a doubling time of approximately 28 hours in RPMI 1640 medium supplemented with 10% fetal bovine serum (FBS). In ACL-3 medium, the doubling time is slightly extended to 38 hours, while in ACL-3 supplemented with bovine serum albumin (BSA), it reaches 42 hours. These variations in doubling time provide valuable insights into cell behaviour under different experimental conditions. At passage 60, A427 cells display a hypotriploid to hypertriploid karyotype. This means the cells possess abnormal chromosomes, including dicentrics, minutes, and a large subtelocentric marker. Such karyotypic abnormalities are often associated with cancer cells and contribute to the unique characteristics of this cell line. A427 cells exhibit tumorigenic properties, allowing them to form tumours when injected into nude mice. These tumours resemble undifferentiated adenocarcinoma, further emphasizing the relevance of this cell line in studying lung cancer and its progression. With its exceptional features, A427 cells find utility in various applications, particularly in cancer research. Their epithelial morphology and lung origin make them an ideal model for studying lung cancer and related diseases. Additionally, A427 cells are well-suited for 3D cell culture techniques, providing a more physiologically relevant environment to explore the behaviour of lung cancer cells.