21/07/2023

A375

A375 is a human melanoma cell line widely used in toxicology and immuno-oncology research. Researchers employ this cell line to study cancer biology, and signalling pathways, test or screen potential anticancer agents, and develop new and effective therapeutics.

This article encompasses all the useful information about A375 melanoma cells you require to start working with it. Here you will know:

Origin and general characteristics of A375 cells
A375 cells: Cell culture information
Advantages & Limitations of A375 cell line
Research applications of A375 cell line
A375 cells: Publications
Resources for A375 cell line: Protocols, Videos, and More

1. Origin and general characteristics of A375 cells

Knowing about a cell line's origin and general characteristics can help you plan its use in research. This section of the article will cover the origin and features of the A375 cell line. For instance, what is the A375 cell line? What are A375 cell line characteristics? Where do A375 cells come from? What is the morphology of A375 melanoma cells? What is the A375 cell size?

The skin cancer cell line A375 originated from the explant culture of a solid tumour of a 54-year-old female with malignant melanoma [1].
A375 cells have epithelial-like morphology.
The A375 cell size is relatively smaller compared to other cell lines. They roughly possess a diameter of 12 µm.
The A375 melanoma cells are hypotriploid. The modal chromosome number for this cell line is 62. Usually, each cell has nine marker chromosomes and one copy of normal N2, N6, and N22 chromosomes.

A doctor examines the growths on an adult's skin for skin cancer.

2. A375 cells: Cell culture information

A375 cell cultures are easy to maintain. They do not have any fussy cell culture requirements. This section will help you learn the necessary cell culture information. Such as what is the A375 doubling time. What is the A375 cell line media? What is the seeding density of A375? Are A375 melanoma cells adherent?

Key Points for Culturing A375 Cells

Population doubling time:	The A375 doubling time is 20 hours.
Adherent or in Suspension:	A375 is an adherent human melanoma cell line.
Seeding Density:	1 x 10⁴ cells/cm²cell density is ideal for A375 cell cultures. Adherent cells are rinsed with 1 x phosphate buffer saline (PBS) and incubated with passaging solution (Accutase). Afterwards, cells were added with culture media and centrifuged. Harvested cells were resuspended in a culture medium and dispensed into new flasks for growth.
Growth Medium:	DMEM is recommended as ideal A375 media when supplemented with 10% fetal bovine serum (FBS), 4.5 g/L Glucose, 1.5 g/L NaHCO3, 4 mM L-Glutamine, and 1.0 mM Sodium pyruvate. A375 media should be renewed 2-3 days per week.
Growth Conditions:	A375 melanoma cells are grown in a humidified incubator (37°C) with a 5% CO2 supply.
Storage:	Frozen A375 cells should be kept below -150°C in the vapour phase of liquid nitrogen or an electric freezer.
Freezing Process and Medium:	CM-1 or CM-ACF media are used to freeze the A375 cell line. A slow freezing process that allows a gradual 1°C decrease in temperature has opted to protect cell viability.
Thawing Process:	The frozen A375 melanoma cells vial is rapidly agitated in the water bath for 40 to 60 seconds until only a small ice clump is left. Cells are added with fresh media and centrifuged to remove freezing media elements. The obtained cell pellet was again resuspended and poured into new flasks.
Biosafety Level:	A375 cultures are handled and maintained in biosafety level 1 laboratory settings.

Confluent layer of adherent A375 cells at 20x magnification.

3. Advantages & Limitations of A375 cell line

Like other cell lines, the A375 melanoma cell line is associated with a unique combination of advantages and limitations, making it a valuable tool for melanoma cancer research. This section has listed only a few significant pros and cons of the A375 cell line.

Advantages

The significant advantages of A375 cells are mentioned below:

Melanoma tumour model	A375 cell line was derived from a malignant melanoma tumour, thus serving as an invaluable tumour model to investigate melanoma biology and therapeutic interventions. These cells exhibit melanoma-like properties, such as a high proliferation rate and invasiveness.
Tumorigenicity	A375 cells are highly tumorigenic, meaning they can form tumours. A375 cells are reported to produce aggressive amelanotic melanomas in NIH Swiss mice treated with anti-thymocyte serum (an immune-suppressive agent).
Well-characterized	A375 is a genetically well-characterized cell line. It possesses specific genetic mutations like melanoma. A375 mutations include A375 BRAF gene alteration, making it sensitive to BRAF-specific inhibitors. This genetic profile thus makes A375 cells a great cancer research tool to study underlying molecular pathways and targeted therapies.

Limitations

The limitation associated with A375 cells are:

In vitro limitations

A375 is an in vitro tumour model. Although it mimics the characteristics of melanoma, it cannot fully represent the complexity of melanoma tumours. Therefore, caution is exercised when findings in vitro are generalized to clinical studies.

4. Research applications of A375 cell line

The A375 cell line has extensive research applications in cancer research. Here are some of the common areas where A375 cells are utilized.

Cancer research: A375 cells have widespread applications in melanoma research. Researchers used these cells to investigate skin cancer biology, processes, and underlying cell signalling mechanisms, including cell proliferation, migration, and invasion. This may also help researchers to identify new molecular therapeutic targets. A study conducted by Lin Zhu and colleagues in 2019 used highly aggressive melanoma cells A375 and found the role of microRNA-3662 on the growth and invasion of A375 melanoma cells. The study findings state that microRNA-3662 targets the ZEB1 gene and regulates melanoma cell growth in vitro and in vivo in the A375 tumour model [2]. Similarly, research conducted in 2018 found that PI3K/AKT and MAPK/ERK pathways are involved in the proliferation and metastasis of the A375 cell line [3].
Drug discovery and testing: A375 cell line is a useful in vitro tumour model to test and screen potential anticancer agents. Researchers evaluate the toxicity and efficacy of novel chemotherapeutic drugs, compounds, and other therapies using these A375 melanoma cells. For example, a study explored the anticancer properties of grapefruit-derived micro and nanovesicles on A375 human melanoma cancer cells. The results revealed that micro and nanovesicles induce cell cycle arrest and apoptosis and hamper gene expression in cell proliferation, migration, and invasion [4].

5. A375 cells: Publications

Some significant research publications featuring A375 cells are mentioned here.

Vitamin D and its low calcemic analogs modulate the anticancer properties of cisplatin and dacarbazine in the human melanoma A375 cell line

This study was published in the Journal of Oncology in 2019. The study proposed that co-treatment with vitamin D and analogues improves the anticancer activity of cisplatin and dacarbazine in A375 melanoma cells.

In vitro evaluation of the anti-melanoma effects (A375 cell line) of the gel and whole leaf extracts from selected aloe species

This research was published in the Journal of Herbal Medicine in 2022. This study explored the anti-melanoma effects of aloe vera gel in A375 melanoma cells.

Fermitin family member 2 promotes melanoma progression by enhancing the binding of p-α-Pix to Rac1 to activate the MAPK pathway

This study in Oncogene (2021) proposed that fermitin family member 2 (FERMT2 or kindlin-2) encourages melanoma progression via activating MAPK through binding with p-α-Pix and Rac1.

FARP1 facilitates cell proliferation through modulating MAPK signaling pathway in cutaneous melanoma

This study in The American Journal of Dermatopathology (2019) proposed that FARP1 may facilitate cutaneous melanoma development and progression. Thus, it can be a useful therapeutic target.

Cytotoxic and apoptotic potential of Phyllodium elegans extracts on human cancer cell lines

This research article was published in Bioengineered (2019). This study explored the cytotoxic and apoptotic activity of a plant Phyllodium elegans extract on human carcinoma cell lines, including A375.

6. Resources for A375 cell line: Protocols, Videos, and More

The following are a few online resources featuring A375 cells.

A375 cell transfection: This video tutorial will help you learn A375 cell transfection step by step.
Cell culturing: This video has very useful information regarding cell line culturing.

The cell culture protocols for A375 cells are listed here.

A375 cell line: This link contains useful information about A375 cell line culturing and maintenance, including A375 media and handling of frozen and proliferating A375 cultures.

References

Avram, S., et al., Standardization of A375 human melanoma models on chicken embryo chorioallantoic membrane and Balb/c nude mice. Oncol Rep, 2017. 38(1): p. 89-99.
Zhu, L., et al., MicroRNA-3662 targets ZEB1 and attenuates the invasion of the highly aggressive melanoma cell line A375. Cancer Manag Res, 2019. 11: p. 5845-5856.
Peng, X., et al., Oxyfadichalcone C inhibits melanoma A375 cell proliferation and metastasis via suppressing PI3K/Akt and MAPK/ERK pathways. Life sciences, 2018. 206: p. 35-44.
Stanly, C., et al., Grapefruit-derived micro and nanovesicles show distinct metabolome profiles and anticancer activities in the A375 human melanoma cell line. Cells, 2020. 9(12): p. 2722.