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CAL27 Cells - Focused Insights into Head and Neck Cancer Using CAL27 Cells

CAL27 is one of the most frequently used oral squamous cell carcinoma cell lines in biomedical research. It is a valuable tool for studying the intricacies of squamous cell cancers, including tumour growth, development, and underlying cellular and molecular mechanisms. Besides, it also facilitates drug screening and testing for anti-cancer drug development.

This article will offer you valuable insights about the CAL27 cell line and will cover the following topics:

  1. Origin and general characteristics of CAL27 cells
  2. Culturing information about CAL27 cell line
  3. CAL27 cell line: Advantages & Disadvantages
  4. Applications of CAL27 cells in research
  5. Publications featuring CAL27 cell line
  6. Resources for CAL27 cell line: Protocols, Videos, and More

1.      Origin and general characteristics of CAL27 cells

The first thing you need to know about a cell line is its origin and general characteristics that distinguish it from other cell lines. This section of the article encompasses primary information about CAL27 cells. You will learn: What is the CAL27 cell model? What is CAL27 HPV? What is CAL27 HER2 status? What is the origin of CAL27 cell line? What is the morphology of CAL27 cells? What is CAL27 cell size?

  • The oral squamous cell cancer cell line, CAL27 was developed from the tongue tissue of a 56-year-old white male presented with tongue squamous cell carcinoma disease. The cell line was established in 1982 by Gioanni and his colleagues [1].
  • The CAL27 cancer cells exhibit an epithelial-like morphology.
  • These cells have an aneuploid karyotype. They possess a modal chromosome number of 43.

An array of squamous epithelial cells observed under the microscope.

2.      Culturing information about CAL27 cell line

For easy and feasible cell line maintenance, you must know its key cell culture requirements. This section covers the important cell culturing concepts about the CAL27 cell line. You should know: What is the doubling time of the cell line CAL27? What is CAL27 cell media? What is the subcultivation ratio of the CAL27 cell line? What are CAL27 culture conditions? How do you culture CAL27 cells?

Key Points for Culturing CAL27 Cells

Doubling Time:

The CAL27 cells exhibit a moderate growth rate with a population doubling time of approximately 45 hours.

Adherent or in Suspension:

These polygonal CAL27 cells adhere to the surface of the culture vessel and grow into monolayers.

Split ratio:

CAL27 cells are sub-cultivated at a split ratio of 1:2 to 1:4. For subculturing, adherent cells are washed with magnesium and calcium-free phosphate buffer saline (1x). Afterwards, the cells are incubated with Accutase (passaging solution) at ambient temperature for 8 to 10 minutes. Detached cells are added with fresh media and centrifuged. The collected pellet was then carefully resuspended and poured into new flasks containing growth media.

Growth Medium:

DMEM media supplemented with 10% FBS, 4.5 g/L Glucose, 4 mM L-Glutamine, 1.5 g/L NaHCO3, and 1.0 mM Sodium pyruvate is used to culture CAL27 cell line. Media should be replaced 2 to 3 times a week.

Growth Conditions:

CAL27 cell cultures are kept at 37°C in a humidified incubator with a 5% CO2 supply.


Frozen CAL27 cells are kept either in the vapor phase of liquid nitrogen or at below -150°C temperature in an ultra-low freezer for longer.

Freezing Process and Medium:

CM-1 or CM-ACF freezing medium is recommended for freezing CAL27 cells. For this, a slow freezing process is recommended that allows only a 1 °C decline in temperature per minute to protect the viability of cells.

Thawing Process:

Frozen cell vials are placed in a water bath pre-set at 37°C for 40-60 seconds until a small ice clump is left. The cells are added with fresh medium and centrifuged. Afterwards, the freezing media is removed, and the cell pellet is added with fresh growth medium and carefully resuspended. The cells are then dispensed into new flasks for growth.

Biosafety Level:

Biosafety level 1 laboratory settings are necessary for CAL27 cell culture handling and maintenance.


Closer look at CAL27 cells at 10x and 20x magnification, highlighting their intricate polygonal structure.

3.      CAL27 cell line: Advantages & Disadvantages

CAL27 is a broadly used oral squamous cell carcinoma cell line. This section will highlight some significant advantages and disadvantages related to these cancer cells.


The main advantages of the CAL27 cell line are:


CAL27 cell line has tumorigenic potential. These cells can develop tumours when injected into nude or immunocompromised mice. This may help researchers understand tumour biology and microenvironment in depth.

Genetic Mutations

CAL27 cells harbour specific genomic alterations, such as CAL27 P53, commonly occurring in oral squamous cell carcinoma. Thus, using CAL27 can be advantageous for researchers studying the impact of these mutations on cancer growth, development, and progression.

Easy to Culture

The oral squamous cell line, CAL27, has no fussy cell culture requirements. Thus, it is easy to culture and maintain in research laboratories.



The disadvantages associated with the CAL27 cell line are:

In Vitro Model

CAL27 cells serve as an in vitro model of oral squamous cell carcinoma. However, it's important to note that they may not accurately reflect the characteristics and complexity of the original squamous cell tumours.


4.      Applications of CAL27 cells in research

CAL27 cell line has a wide range of applications in biomedical research. Some of the important research uses are listed here.

  • Cancer Biology: CAL27 cells have been widely used to explore the molecular and cellular mechanisms underlying the development and progression of oral squamous cell carcinoma (OSCC). Researchers also study genetic mutations, signalling pathways, and important biomarkers related to squamous cell tumours. A study conducted in 2021 utilized CAL27 cells and investigated the role of CD147 in the development of oral squamous cell cancer. The study outcomes showed that CD147 knockdown caused inhibition of CAL27 cell proliferation, migration, and invasion. Besides, its downregulation also impedes the development of drug resistance in cancer cells [2].
  • Drug Development: CAL27 cell line serves as a valuable tool for screening and testing of anti-tumor drugs. Researchers employ these cells to test the efficacy of various chemotherapeutic agents, targeted therapies, and experimental drugs in inhibiting oral cancer cells' proliferation, growth, and viability. Moreover, the modulating effect of the potential anti-cancer agents has also been assessed on the dysregulated cancer cell signalling pathways. This may assist in developing effective and personalized treatments to fight disease. A study explored the anti-tumor potential of zinc oxide nanoparticles using CAL27 cells. The study findings revealed that zinc oxide nanoparticles produced cytotoxic effects and also induced the PINK1/Parkin pathway-mediated mitophagy in CAL27 tongue cancer cells [3]. Similarly, researchers observed the anti-cancer potential of doxorubicin and its nano-formulated form (Doxil) on CAL27 oral squamous cell cancer [4].

5.      Publications featuring CAL27 cell line

This section of the article will cover a few interesting and most cited publications on CAL27 cancer cells:

Inhibition of CAL27 oral squamous carcinoma cell by targeting hedgehog pathway with vismodegib or itraconazole

This research was published in the Frontiers in Oncology (2020). The study explored the anti-oral squamous cell cancer effects of vismodegib or itraconazole using CAL27 cells.  

PLAU promotes cell proliferation and epithelial-mesenchymal transition in head and neck squamous cell carcinoma

This article in Frontiers in Genetics (2021) proposed that plasminogen activator, urokinase (PLAU) protein, encourages the proliferation, migration, and epithelial to mesenchymal transition (EMT) of CAL27 cells.

MicroRNA-145-5p aggravates cell apoptosis and oxidative stress in tongue squamous cell carcinoma

This article in the Experimental and Therapeutic Medicine (2021) found that miRNA-145-5p aggravates CAL27 cells' oxidative stress and death and suppresses the PI3K/AKT signalling pathway.

Antitumor activity of the plant extract morin in tongue squamous cell carcinoma cells

This study in Oncology Reports (2018) explored the anti-cancer activity of a bioflavonoid called morin, utilizing CAL27 cells.

Reduction of oral pathogens and oxidative damage in the CAL 27 cell line by Rosmarinus officinalis L. and Taraxacum officinale Web. Extracts

This article in the Journal of Ethnopharmacology (2023) investigated the therapeutic potential of Rosmarinus officinalis L. and Taraxacum officinale extracts on CAL27 cells.

6.      Resources for CAL27 cell line: Protocols, Videos, and More

There are limited available resources on the CAL27 cell line. In this section, we have listed some general protocols that can also help you deal with CAL27 cells:

The following link contains the CAL27 cell culture protocol.

  • CAL27 cell line: This website contains information about CAL27 media, seeding density, doubling time, etc. Besides, it contains a protocol for subculturing and handling proliferating and cryopreserved cultures.


  1. Liu, X., et al., LINC00472 suppresses oral squamous cell carcinoma growth by targeting miR-455-3p/ELF3 axis. Bioengineered, 2022. 13(1): p. 1162-1173.
  2. Pan, S., et al., Knockout of CD147 inhibits the proliferation, invasion, and drug resistance of human oral cancer CAL27 cells in Vitro and in Vivo. International Journal of Biological Macromolecules, 2021. 181: p. 378-389.
  3. Wang, J., et al., Zinc oxide nanoparticles induce toxicity in CAL 27 oral cancer cell lines by activating PINK1/Parkin-mediated mitophagy. International Journal of Nanomedicine, 2018: p. 3441-3450.
  4. Abd El-Hamid, E.S., A.M. Gamal-Eldeen, and A.M.S. Eldeen, Liposome-coated nano doxorubicin induces apoptosis on oral squamous cell carcinoma CAL-27 cells. Archives of oral biology, 2019. 103: p. 47-54.