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Cell Line Profile A549 

(ECACC Catalogue no. 86012804)

Purchase Cell line


Cell line history

The A549 cell line was isolated in 1973 from a pulmonary adenocarcinoma1 in a study to attempt to establish continuous cell lines from 200 different tumours. Despite being developed as a tool for cancer research the cell line was consequently characterized as being representative of the Alveolar Type II pneumocytes of the human lung 2 3 4 5 and because of this the cell line has been a mainstay of respiratory research for nearly forty years. Work with early passage A549 soon after the isolation of the cell line provided evidence of its ability to exhibit features of an ATII epithelial cell phenotype3 4 5, together with reports that extended culture of early passage material resulted in cellular ‘differentiation’ evident by the expression of high numbers of Multilamellar bodies (MLB) 2 6 indicating the cell might be capable of surfactant production, a key feature of ATII cells. More contemporary studies7 8, however, have led to a more conservative view on the cell line’s suitability to recapitulate the ATII phenotype. Recent work9 shown that extended culture in physiologically relevant cell culture medium may increase the ATII like properties of the cell line.

A459 monolayer stained for lipid droplets using Oil-Red O

Transmission Electron Micrograph (TEM) of Multilamellar bodies (MLB) in an A549 cell after long term culture (n=nucleus).

Key characteristics

Using the cytidine diphosphocholine pathway the cells can synthesise phospholipids (lecithins) which manifest as lipid droplets in the cytoplasm. With long term culture in a physiological medium the lipid may be expressed as Multilamellar Bodies (MLB) similar to those seen in ATII pneumocytes. Extensive RNA microarray analysis of the gene expression of the cell line is openly available9. 


Cancer research, the generation of epithelial models of the distal lung for studies into airway function and disease and virology; the A549 cell line is a suitable host for many human respiratory viruses including adenoviruses.

Culture tips

Cultures of the cell line can be expanded effectively using conventional subculture techniques, however, for lung disease research it is recommended that the cells are maintained in long term culture (around 20 days) with regular medium changing with a physiologically relevant medium such as Hams F12/10% FBS to allow differentiation to an ATII like phenotype. Culture in simple medium such as DMEM/10% FBS may result in the adoption of a mucus secreting phenotype not typical of distal lung.

Related cell lines

ECACC catalogue number




Human Bronchial Epithelium



Human lung epidermoid carcinoma

Key references

1. Giard, D. J. et al. In vitro cultivation of human tumors: establishment of cell lines derived from a series of solid tumors. J. Natl. Cancer Inst. 51, 1417–1423 (1973)

2. Shapiro, D. L., Nardone, L. L., Rooney, S. A., Motoyama, E. K. & Munoz, J. L. Phospholipid biosynthesis and secretion by a cell line (A549) which resembles type II aleveolar epithelial cells. Biochim. Biophys. Acta 530, 197–207 (1978)

3. Lieber, M., Smith, B., Szakal, A., Nelson-Rees, W. & Todaro, G. A continuous tumor-cell line from a human lung carcinoma with properties of type II alveolar epithelial cells. Int. J. Cancer J. Int. Cancer 17, 62–70 (1976)

4. Foster, K. A., Oster, C. G., Mayer, M. M., Avery, M. L. & Audus, K. L. Characterization of the A549 Cell Line as a Type II Pulmonary Epithelial Cell Model for Drug Metabolism. Exp. Cell Res. 243, 359–366 (1998)

5. Nardone, L. L. & Andrews, S. B. Cell line A549 as a model of the type II pneumocyte: Phospholipid biosynthesis from native and organometallic precursors. Biochim. Biophys. Acta BBA - Lipids Lipid Metab. 573, 276–295 (1979)

6. Balis, J. U., Bumgarner, S. D., Paciga, J. E., Paterson, J. F. & Shelley, S. A. Synthesis of lung surfactant-associated glycoproteins by A549 cells: description of an in vitro model for human type II cell dysfunction. Exp. Lung Res. 6, 197–213 (1984)

7. Swain, R. J., Kemp, S. J., Goldstraw, P., Tetley, T. D. & Stevens, M. M. Assessment of Cell Line Models of Primary Human Cells by Raman Spectral Phenotyping. Biophys. J. 98, 1703– 1711 (2010)

8. Corbière, V. et al. Phenotypic characteristics of human type II alveolar epithelial cells suitable for antigen presentation to T lymphocytes. Respir. Res. 12, 15 (2011)

9. Cooper, J. R. et al. Long Term Culture of the A549 Cancer Cell Line Promotes Multilamellar Body Formation and Differentiation towards an Alveolar Type II Pneumocyte Phenotype. PLOS ONE 11, e0164438 (2016)