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A549 Cells - Comprehensive Guide to A549 Cell Line for Pulmonary Cancer Research

The A549 cell line, a non-small cell lung cancer cell line, is derived from human adenocarcinoma alveolar epithelial cells. These cells have a broad range of research applications in cancer, toxicology screening, and immuno-oncology research. In this article, we will provide all the necessary information for working with this cell line, including the origin and general characteristics of A549 cells.

  1. A549 Origin and General Information
  2. A549 and its Applications in Research
  3. Culture Information for A549 cells
  4. Pros and Cons of the A549 Cell Line: What You Need to Know
  5. A549 cells: Research Publications
  6. Resources for A549: Protocols, Videos, and Other Useful Materials

Overall, this article will provide a comprehensive understanding of the A549 cell line to facilitate its use in various research fields.

1.       A549 Origin and General Information

A549 cells were isolated from the lung cancer tissue of a 58-year-old male (Caucasian) in 1972, and established as a cell line by D. J. Giard and colleagues. A549 cells are adherent and grow in monolayers, and have an epithelial-like morphology that is similar to squamous cells found in lung tissue.

The mean cell diameter for the A549 cell line ranges from 10.59µm to 14.93µm, which is determined by using transmission electron microscope and inverted microscope. A549 cells are hypotriploid, which means they have fewer chromosomes than human cells. The modal chromosome number for 24% of cells is 66, while higher ploidies occur infrequently (0.4%) with 64 or 67 chromosome numbers.

2.       A549 and its Applications in Research

A459 is an FDA-approved cell line that has many applications in the molecular biology field. It is used to produce therapeutic constructs evaluated in clinical trials. Important research applications of A549 are:

Production of Adenovirus

A549 is a viable source for producing adenoviruses, as these viral constructs do not require additional viral oncogene (E1A or early region 1A) for the transcription of the viral gene. Adenoviral-based therapies and vaccine strategies use these constructs in clinical trials. Additionally, the A549 cell line is used in assays as a control to measure the replication of adenovirus constructs deficient in the E1A gene [1].

Drug Screening and Development

The A549 cell line is a valuable model for drug screening, as demonstrated in recent research. For example, in 2021, a study used A549 cells to screen 65 plants collected from Western Andalusia, Spain for selective anticancer activity [2]. Additionally, another study explored the anti-cancer potential of green nanoparticles using the A549 cell line [3].

A549 as a Disease Model

Due to their origin, the A549 lung carcinoma-derived epithelial cells have become a popular disease model for lung cancer research [4]. For instance, a recent study employed A549 cells to develop a cell line-based xenograft model of lung carcinoma for investigating tumor growth and metastasis [5]. Additionally, the A549 cell line is a reliable mimic of type II alveolar epithelium [6]. This makes it a valuable tool for studying metabolic processes in lung tissue and investigating drug delivery to the pulmonary tissue.

3.       Culture Information for A549 cells

In the below table, we have summarized all the information required to work with the A549 cell line. This includes the population doubling time, adherent growth characteristics, recommended seeding density, growth medium, and more.



Population doubling time

28 hours

Adherent or in suspension

Adherent cell line that grows in monolayers

Seeding density

Recommended seeding density is 1 x 104 cells/cm2

Growth medium

DMEM or Ham's F12 growth medium with 5% FBS and L-glutamine

Growth conditions

Humidified incubator at 37°C with 5% CO2


Liquid nitrogen vapour phase

Freezing process and medium

Slow freezing process with CM-ACF or CM-1 freezing media

Thawing process

Rapid agitation in a pre-warmed water bath at 37°C, followed by centrifugation and culturing in a flask with fresh medium

Biosafety level

Biosafety level 1

4.      Pros and Cons of the A549 Cell Line: What You Need to Know

The A549 cell line is a popular choice for researchers studying lung cancer, but it is important to understand both its advantages and limitations before use. On one hand, A549 cells are a well-characterized cellular model for lung cancer research and have easy-to-follow culturing requirements. On the other hand, A549 cells are susceptible to microbial contamination like other cell lines and require aseptic culturing conditions to avoid it. Bacterial infections can be detected through pH change and turbidity of the growth medium, but mycoplasma infections can spread rapidly and damage cell morphology and growth rate. Additionally, A549 cell health can deteriorate with extended culture periods, so their passage number should be kept below 20 for optimal cell health. By considering both the advantages and limitations, researchers can make informed decisions when selecting the A549 cell line for their studies.

5.       A549 cells: Research Publications

In this section, you can explore a selection of notable publications on A549 lung carcinoma cells that may inspire new research ideas involving this cell line.

  1. A study conducted in 2020 aimed to develop suspension cultures of A549 cells to enhance the production of oncolytic adenoviruses. These viruses are used for cancer therapy.
  2. This study used the A549 cell line to evaluate the anti-cancer potential of Hypnea valentiae seaweed-loaded green nanoparticles.
  3. This publication in the Bioengineered journal proposed C-met specific chimeric antigen receptor T cell as direct therapy for A549 non-small cell lung carcinoma cells.
  4. Eul-Bee Ko and colleagues published this study in 2022, in which they used A549 cells for anti-cancer drug screening. This study investigated the role of gallic acid, a natural compound, in inducing cell death and preventing lung cancer progression.
  5. This research paper utilized the A549 cell line to validate the anti-tumor activity of combined paclitaxel and curcumin compound nanoformulations.
  6. This publication in Cancer Management and Research journal employed A549 cells as a non-small lung cancer model and study the mechanism behind cisplatin resistance.
  7. This study in Viruses journal explored the varying effect of human adenovirus early region (E1A) protein isoforms on aerobic glycolysis by using the A549 cell line as a cell model.

6.       Resources for A549: Protocols, Videos, and Other Useful Materials

A549 is a well-known lung carcinoma epithelial cell line. There are many informative resources about these cells’ culturing rerquirements, transfection, and much more. Our optimized online protocol will help you learn all about culturing A549 including growth medium, subculturing procedure, freezing and thawing of cells, etc.

A549 Transfection protocols

Transfection protocol: This site provides all necessary information for transfecting A549 cells.

Transfection of A549 cells: This link will lead to DNA transfection protocol for the A549 cell line.

Transfection protocol: A general DNA transfection protocol used for mammalian cell lines is shown in this video.

Videos about the A549 cell line

Cell culture protocol for A549 cells: This video is on culturing of this human lung carcinoma cell line.

A549 general information: This video provides some basic information about the A549 cell line.

With this comprehensive guide to the A549 cell line, you now have all the necessary information to use this powerful tool in your research. Whether you're exploring cancer, toxicology, or immuno-oncology, A549 cells offer a reliable and well-characterized model. If you're ready to get started, don't hesitate to reach out to us or order from our website today. We can't wait to see the groundbreaking discoveries that you'll uncover with A549 cells.


  1. Moreira, A.S., et al., Establishing suspension cell cultures for improved manufacturing of oncolytic adenovirus. Biotechnology Journal, 2020. 15(4): p. 1900411.
  2. Calderón-Montaño, J.M., et al., Screening for selective anticancer activity of 65 extracts of plants collected in Western Andalusia, Spain. Plants, 2021. 10(10): p. 2193.
  3. Viswanathan, S., et al., Anti-cancer activity of Hypnea valentiae seaweed loaded gold nanoparticles through EMT signaling pathway in A549 cells. Biochemical Systematics and Ecology, 2023. 107: p. 104606.
  4. Garcia-de-Alba, C., Repurposing A549 adenocarcinoma cells: New options for drug discovery. 2021, American Thoracic Society. p. 405-406.
  5. Qi, L., et al., In vivo tumor growth and spontaneous metastasis assays using A549 lung cancer cells. Bio-protocol, 2020. 10(7): p. e3579-e3579.
  6. Khan, P., et al., Culture of human alveolar epithelial type II cells by sprouting. Respiratory research, 2018. 19(1): p. 1-7.

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