a H&E and immunohistochemical staining images of lung cancer tissues and derived organoids. Shown are representative examples of LAC with acinar (LC102 and LC98) or solid (LC96) subtypes and LSC (LC97). The LCOs retained the tumor cell organizations and the expression patterns of the characteristic markers (TTF-1 and CK7 for LAC, p40, and CK5/6 for LSC). Scale bars, 20 µm. The experiments are repeated three times. b Heat map illustrating genome-wide copy number variations (CNVs) of lung cancer tissue–LCO pairs. DNA copy number gains (red) and losses (blue) found in the original lung cancer tissues are conserved in the corresponding tumor organoids. Signal amplification can be seen in all the LCOs compared to the original tumor tissues (T tissue, O organoids). c Overview of somatic mutations in cancer genes found in the tissue-organoid pairs. Shown is the most severe mutation per gene. d Stacked bar graphs illustrating the numbers of passages and the freeze–thaw status of 20 LCO lines underwent long-term culture. Each block indicates one passage or a freeze–thaw cycle. For example, LC96-O and LC97-O have been passaged more than ten times and successfully thawed after frozen at passage 10. LC116-O has been passaged five times but failed to recover after frozen. LC105-O stopped growing at passage 2. e Bright-field images showing the unchanged morphologies of LC96-O and LC97-O at P0, P3, P5, and P10. Scale bar, 200 µm. The experiments are repeated three times. f Heat map showing the increasing purities of tumor cells with passaging in LC96-O and LC97-O.