| Field | Specification |
|---|---|
| Species |
The A375 human melanoma cell line, isolated from the skin of a 54-year-old female patient with malignant melanoma, is a significant resource in cancer research, particularly in the study of human melanoma, one of the most aggressive forms of skin cancer. The A375 cell line is known for its rapid growth rate and high tumorigenic potential, making them suitable for various experimental applications, including in vitro studies on cell proliferation, migration, and invasion, as well as in vivo tumorigenesis assays.
A375 cells exhibit high tumorigenic potential in immunosuppressed mice, forming rapidly growing amelanotic melanomas. The presence of the BRAFV600E mutation in A375 cells makes them highly sensitive to MEK inhibition, providing a valuable tool for investigating targeted therapies in melanoma treatment. The treatment of A375 cells with vemurafenib, for example, has been shown to enhance the induction of MHC Class I and Class II molecules, offering insights into the interactions between melanoma cells and the immune system.
In addition to their role in basic melanoma research, A375 cells are used in drug screening and in the investigation of signaling pathways involved in cancer cell survival, proliferation, and metastasis. A375 cells have further been utilized in apoptosis studies and A375 isogenic cell lines and the introduction of reporter proteins like luciferase (luc2) enable the study of gene function and the monitoring of cellular responses in real-time. A375 cells' suitability as a transfection host and their use in stable reporter cell lines also contribute to their versatility in research applications.
In summary, the A375 human melanoma cell line is a pivotal tool in the investigation of human melanoma, offering a comprehensive model for studying the molecular and cellular mechanisms underlying melanoma progression, the efficacy of therapeutic agents, and the interaction between cancer cells and the immune system.
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- Antigen expression: P53 positive
- Tumorigenic: Yes, in nude mice
- Mutational profile: BRAF V600Emut
- Karyotype: A375 cells are characterized by their hypotriploid karyotype, with a modal chromosome number of 62, and the presence of nine marker chromosomes in each cell, highlighting the genetic alterations associated with malignant melanoma.
- cultureMedium: DMEM, w: 4.5 g/L Glucose, w: 4 mM L-Glutamine, w: 3.7 g/L NaHCO3, w: 1.0 mM Sodium pyruvate (Cytion article number 820300a)
- supplements: Supplement the medium with 10% FBS
- dissociationReagent: Accutase
- doublingTime: 20 hours
- subculturing: Remove the old medium from the adherent cells and wash them with PBS that lacks calcium and magnesium. For T25 flasks, use 3-5 ml of PBS, and for T75 flasks, use 5-10 ml. Then, cover the cells completely with Accutase, using 1-2 ml for T25 flasks and 2.5 ml for T75 flasks. Let the cells incubate at room temperature for 8-10 minutes to detach them. After incubation, gently mix the cells with 10 ml of medium to resuspend them, then centrifuge at 300xg for 3 minutes. Discard the supernatant, resuspend the cells in fresh medium, and transfer them into new flasks that already contain fresh medium.
- seedingDensity: 1 x 104 cells/cm2 will result in a confluent monolayer within 4 days.
- fluidRenewal: 2 to 3 times per week
- postThawRecovery: After thawing, plate the cells at 4 x 104 cells/cm2 and allow the cells to recover from the freezing process and to adhere for at least 24 hours.
- freezeMedium: As a cryopreservation medium, use complete growth medium (including FBS) + 10% DMSO for adequate post-thaw viability, or CM-1 (Cytion catalog number 800100), which includes optimized osmoprotectants and metabolic stabilizers to enhance recovery and reduce cryo-induced stress.
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