B16-F10 cells constitute a melanoma cell line driven from C57BL/6J mouse. They are widely used in skin cancer research. Researchers use these cells to study tumour development and progression and therapeutic interventions. This article will cover the fundamental aspects of B16-F10 melanoma cells. Notably, it will include:

  1. Origin and general characteristics of the B16-F10 cell line
  2. Culturing information of B16-F10 cells
  3. B16-F10 cells: Advantages & disadvantages
  4. Research applications of B16-F10 cells
  5. Publications featuring B16-F10 Cell line
  6. Resources for B16-F10 Cell line: Protocols, Videos, and More

1.      Origin and general characteristics of the B16-F10 cell line

This section will give you insights into the origin and distinctive attributes of B16F10 melanoma tumour cells. It will help you use the cell line efficiently in your research work. Mainly, you will learn: What are B16-F10 cells? What is B16F10 derived from? What is the morphology B16F12 cell line? What is the size of the B16F10 cell?

  • B16-F10 is a subclone of the B16 tumour cell line derived from C57BL/6J mice skin tissue. Herein, B16F10 melanoma cells were developed after intravenously injecting the B16 line into immunocompromised or syngeneic mice. These cells were selected for their potential to form metastasized lung colonies in vivo and then established after ten cycles of lung colony formation in vitro [1]. It was developed by Fidler and colleagues in 1976.
  • B16-F10 cell lines have an epithelial-like and spindle-shaped appearance.
  • The approximate size of B16-F10 cells is 15.4 ± 1.4 μm [2].

B16-F1 and B16-F10 cells

B16-F1 and B16-F10 cells were derived from the B16 parent cell line. Both originated in the same and possess almost similar characteristics. However, the main difference is their metastatic ability. B16-F10 cells have high, whereas B16-F1 has low metastatic potential [3].

Highly magnified crosssection of a malignant melanoma tumour under the microscope.

2.      Culturing information of B16-F10 cells

Before handling and cultivating a cell line, you must know about its doubling time, growth media, conditions, and cell culture protocols. This section will discuss: What is the doubling time of b16-f10 cells? How do you culture B16F10 cells? What is the B16-F10 cell media? What culture conditions are recommended for B16-F10 cells?

Key Points for Culturing B16-F10 Cells

Doubling Time:

The doubling time of B16-F10 cells is approximately 20.1 hours. It may range from 17 to 21 hours, depending upon culturing conditions.

Adherent or in Suspension:

B16-F10 is an adherent cell line. The cells grow fast and make monolayers.

Split ratio:

B16-F10 cells are sub-cultivated at a split ratio of 1:2 to 1:4. Cells are washed with phosphate buffer saline (1x) and then incubated with Accutase passaging solution for 8 to 10 minutes at ambient temperature. Cells are added with fresh medium and centrifuged. The harvested cell pellet is again resuspended, and cells are dispended into the new flask containing fresh culture media according to split ratio.

Growth Medium:

B16-F10 cells are cultured in the DMEM medium. The media is supplemented with 10% FBS, 4 mM L-Glutamine, 1.5 g/L NaHCO3, 4.5 g/L Glucose, and 1.0 mM Sodium pyruvate for ideal cell growth. Media should be replaced 2 to 3 times per week.

Growth Condition:

B16-F10 cells are grown in a humidified incubator at 37 °C with a 5% CO2 supply.


Frozen cells are stored below -150 °C in an electric ultra-low temperature freezer or in the vapour phase of liquid nitrogen to maintain cell viability.

Freezing Process and Medium:

B16-F10 cells are frozen in CM-1 or CM-ACF media for storage. For this, a slow freezing process that allows only a 1°C decrease in temperature per minute is recommended to prevent cells from any shock.

Thawing Process:

Frozen B16-F10 cells are thawed in a pre-set 37°C water bath for 40 to 60 seconds. Next, the cells are added to fresh medium and centrifuged to remove freezing media components. The collected cells are resuspended in a growth medium and poured into flasks for culturing.

Biosafety Level:

Biosafety level 1 laboratory is required to handle and maintain the B16-F10 cell line.

Semi-confluent B16-F10 cells at 20x and 10x magnification.

3.      B16-F10 cells: Advantages & disadvantages

Like other cell lines, B16-F10 also exhibits some advantages and disadvantages. Some significant pros and cons of this skin melanoma cell line are discussed in this section.


The B16-F10 cell line is widely used in cancer research. The advantages of B16-F10 cells are:

Metastatic potential

Skin melanoma B16F10 cells exhibit high metastatic potential, making them valuable for studying cancer metastasis and underlying mechanisms.

In vitro tumour model

B16-F10 cells serve as an in vitro model for studying cancer progression and growth, helping researchers understand the cellular and molecular mechanisms driving cancer.



The disadvantages associated with the B16-F10 cell line are:

Mouse-derived cell line

B16-F10 is a mouse-derived cell line, limiting its applicability to human-specific studies. Research findings from these cells may not always truly translate to human biology.


4.      Research applications of B16-F10 cells

The B16-F10 cell line is extensively used in cancer research. A few promising applications of this cell line are discussed here.

  • Cancer research: The B16-F10 cell line is a valuable model to study cancer cell processes, including proliferation, invasion, migration, and cell death or apoptosis. Besides, it helps researchers to gain insights into molecular mechanisms and pathways driving these cellular processes. A study conducted in 2018 explored the role of CCR5 (C-C chemokine receptor type five) in melanoma cells epithelial to mesenchymal cell transition and metastasis. The findings revealed that CCR5 deficiency restricts tumour growth and metastasis, whereas high expression leads to enhanced growth and metastasis of B16-F10 cells. Further research reported that CCR5 regulates TGFβ1 expression, which regulates PI3K/AKT/GSK3β signalling to promote the epithelial to mesenchymal transition and cell migration [4].
  • Drug testing and development: B16F10 melanoma tumor cells are highly aggressive and thus suitable for testing potential anti-tumour drugs and treatments. Researchers employ these cells and assess the effect of different compounds on cell growth, proliferation, and metastasis, aiding drug development. A study carried out in 2018 by Valentina Nanni and colleagues investigated the therapeutic effects of Spartium junceum flowers hydroalcoholic extract. The study proposed that flower extract was effective in inducing senescence in B16-F10 cells, which leads to cell growth and melanogenesis suppression thus, it can exert potential anti-cancer activities [5].

5.      Publications featuring B16-F10 Cell line

Here are some significant research publications featuring the B16-F10 melanoma cell line:

Anti-Melanogenic Effect of Ethanolic Extract of Sorghum bicolor on IBMX–Induced Melanogenesis in B16/F10 Melanoma Cells

This study was published in Nutrients (2020). It proposed that Sorghum bicolor ethanolic extract has an anti melanogenic effect in skin melanoma B16F10 cells.

Calcitriol Inhibits Proliferation and Potentially Induces Apoptosis in B16–F10 Cells

The research published in Medical Science Monitor Basic Research (2022) proposed that calcitriol drug exert anti-tumor effects in B16-F10 melanoma cells by inhibiting proliferation and inducing apoptosis.

Prooxidative effect of cardols is involved in their cytotoxic activity against murine B16–F10 melanoma cells

This article is published in Biochemical and Biophysical Research Communications (2022). The findings revealed that Cardols, resorcinolic lipids, exert intense cytotoxicity on the B16-F10 cell line.

Ginkgo biloba exocarp extract inhibits the metastasis of B16-F10 melanoma involving PI3K/akt/NF-κB/MMP-9 signaling pathway

The study published in Evidence-Based Complementary and Alternative Medicine (2018) explored the antimetastatic potential of Ginkgo biloba exocarp extract using B16-F10 cells.

Thymoquinone induces apoptosis in B16-F10 melanoma cell through inhibition of p-STAT3 and inhibits tumor growth in a murine intracerebral melanoma …

This research in World Neurosurgery (2018) proposed that thymoquinone can be an effective therapy against intracerebral metastatic lesions as it suppresses B16-F10 cell growth and induces apoptosis. 

6.      Resources for B16-F10 Cell line: Protocols, Videos, and More

B16F10 endothelial cells are widely used in skin cancer research. Here are some online resources explaining its culturing and transfection protocols:

The following link contains the cell culture protocol for B16-F10 cells:

  • B16-F10 subculturing: This website contains helpful information on B16F10 melanoma tumour cells. It includes growth media, doubling time, culture conditions and protocol for subculturing cells, and handling cryopreserved and proliferative cultures.


  1. Poste, G., et al., Comparison of the metastatic properties of B16 melanoma clones isolated from cultured cell lines, subcutaneous tumors, and individual lung metastases. Cancer Research, 1982. 42(7): p. 2770-2778.
  2. Nakamura, M., D. Ono, and S. Sugita, Mechanophenotyping of B16 Melanoma Cell Variants for the Assessment of the Efficacy of (-)-Epigallocatechin Gallate Treatment Using a Tapered Microfluidic Device. Micromachines, 2019. 10(3): p. 207.
  3. Danciu, C., et al., Behaviour of four different B16 murine melanoma cell sublines: C57BL/6J skin. Int J Exp Pathol, 2015. 96(2): p. 73-80.
  4. Liu, J., et al., High expression of CCR5 in melanoma enhances epithelial–mesenchymal transition and metastasis via TGFβ1. The Journal of Pathology, 2019. 247(4): p. 481-493.
  5. Nanni, V., et al., Hydroalcoholic extract of Spartium junceum L. flowers inhibits growth and melanogenesis in B16-F10 cells by inducing senescence. Phytomedicine, 2018. 46: p. 1-10.