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Updated: Jul 1, 2026
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Generation of 3D Whole Lung Organoids from Induced Pluripotent Stem Cells for Modeling Lung Developmental Biology and Disease
Published on: April 12, 2021
Generating Tumor Organoid Cultures from the Lung Adenocarcinoma Cell Line
Xinzhou Zhao1, Yuanlong He1, Xinhua Lin2
1State Key Laboratory of Genetics and Development of Complex Phenotypes, School of Life Sciences, Fudan University.
Journal of Visualized Experiments : Jove
|June 29, 2026
Summary
We developed a simple, cost-effective protocol for creating 3D lung adenocarcinoma organoids from A549 cells. This Cell Line-Derived Tumor Organoid (CDTO) model is ideal for research and teaching 3D tumor culture techniques.
Area of Science:
- Biomedical Engineering
- Cancer Research
- Cell Biology
Background:
- Lung adenocarcinoma is a major cause of cancer mortality.
- 3D organoid models offer a more physiologically relevant platform for studying cancer compared to 2D cultures.
- Developing robust and reproducible organoid culture protocols is crucial for advancing cancer research.
Purpose of the Study:
- To describe a protocol for generating Cell Line-Derived Tumor Organoids (CDTOs) from the A549 human lung adenocarcinoma cell line.
- To characterize the generated CDTOs for their suitability in lung adenocarcinoma research.
- To present a cost-effective and straightforward organoid platform for both research and educational purposes.
Main Methods:
- Embedding 2D-expanded A549 cells in Matrigel for 3D culture.
- Utilizing a specific seeding density (500 cells/µL) for consistent organoid formation.
- Characterizing CDTOs using hematoxylin and eosin (H&E) staining and immunofluorescence (IF).
Main Results:
- The protocol successfully generated CDTOs from A549 cells.
- Organoids maintained high expression of lung adenocarcinoma markers: TTF-1, ECAD, and KRT7.
- Dysregulated polarity was observed via Zona Occludens 1 (ZO-1) expression, indicating tumor characteristics.
Conclusions:
- The described protocol provides a technically straightforward, cost-effective, and reproducible 3D organoid platform for lung adenocarcinoma research.
- The CDTO model is suitable for studying tumor biology and evaluating therapeutic strategies.
- The protocol's simplicity makes it valuable for undergraduate laboratory teaching of 3D tumor organoid culture techniques.
