A375

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-Luc (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.

37°C, 5% CO₂, humidified atmosphere.

1. Confirm that the vial remains deeply frozen upon delivery, as cells are shipped on dry ice to maintain optimal temperatures during transit.

2. Upon receipt, either store the cryovial immediately at temperatures below -150°C to ensure the preservation of cellular integrity, or proceed to step 3 if immediate culturing is required.

3. For immediate culturing, swiftly thaw the vial by immersing it in a 37°C water bath with clean water and an antimicrobial agent, agitating gently for 40-60 seconds until a small ice clump remains.

4. Perform all subsequent steps under sterile conditions in a flow hood, disinfecting the cryovial with 70% ethanol before opening.

5. Carefully open the disinfected vial and transfer the cell suspension into a 15 ml centrifuge tube containing 8 ml of room-temperature culture medium, mixing gently.

6. Centrifuge the mixture at 300 x g for 3 minutes to separate the cells and carefully discard the supernatant containing residual freezing medium.

7. Gently resuspend the cell pellet in 10 ml of fresh culture medium. For adherent cells, divide the suspension between two T25 culture flasks; for suspension cultures, transfer all the medium into one T25 flask to promote effective cell interaction and growth.

8. Adhere to established subculture protocols for continued growth and maintenance of the cell line, ensuring reliable experimental outcomes.

Mycoplasma contamination is excluded using both PCR-based assays and luminescence-based mycoplasma detection methods.

To ensure there is no bacterial, fungal, or yeast contamination, cell cultures are subjected to daily visual inspections.

For long-term preservation, place vials in vapor-phase liquid nitrogen at about −150 to −196 °C. Storage at −80 °C is acceptable only as a short interim step before transfer to liquid nitrogen.

Cryopreserved cell lines are shipped on dry ice in validated, insulated packaging with sufficient refrigerant to maintain approximately −78 °C throughout transit. On receipt, inspect the container immediately and transfer vials without delay to appropriate storage.

At Cytion, we stand behind the quality and viability of our products. We guarantee that our cells will be viable at the start of culture for 60 days after shipment from our facilities. We assure that all materials conform to the specifications described in the Certificate of Analysis and/or Product Sheet.

For full details, please refer to our Warranty Statement.