A375 Cells

Categories: Skin cancer cell lines Description: 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. Organism: Human Tissue: Skin Disease: Melanoma Synonyms: A 375, A-375, A375-MEL, A375-mel, A375mel Age: 54 years Gender: Female Morphology: Epithelial-like Growth Properties: Adherent Citation: A375 (Cytion catalog number 300110) Biosafety Level: 1 Ncbi_ Taxid: 9606.0 Cellosaurus Accession: CVCL_0132 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. Culture Medium: 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 Dissociation Reagent: Accutase Doubling Time: 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. Seeding Density: 1 x 104 cells/cm2 will result in a confluent monolayer within 4 days. Fluid Renewal: 2 to 3 times per week Post Thaw Recovery: 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. Freeze Medium: 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. Thawing And Culturing Cells: Confirm that the vial remains deeply frozen upon delivery, as cells are shipped on dry ice to maintain optimal temperatures during transit. 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. 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. Perform all subsequent steps under sterile conditions in a flow hood, disinfecting the cryovial with 70% ethanol before opening. 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. Centrifuge the mixture at 300 x g for 3 minutes to separate the cells and carefully discard the supernatant containing residual freezing medium. 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. Adhere to established subculture protocols for continued growth and maintenance of the cell line, ensuring reliable experimental outcomes. Sterility: 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. Safety Precautions: When planning to store a cryovial in liquid nitrogen for future thawing, it is mandatory to adhere to stringent safety measures. Appropriate protective gloves and clothing are essential, and the use of a face mask or safety goggles is required during the transfer of frozen samples to or from the liquid nitrogen tank. This is to mitigate the risk of injury from potential cryovial explosions upon removal, which can result in the projection of sharp fragments. Warranty: We stand by the promise of delivering products with high cell viability and robust culture performance. To achieve the best results, please make sure you follow the storage and culture instructions detailed in the product information sheet closely. Your adherence to these guidelines is key to success. Subject To Material Transfer Agreements: If you intend to use Cytion cell lines solely for internal research at a single research site, please complete and sign our Material Transfer Agreement (MTA) and submit it along with your order.For any commercial applications - including but not limited to fee-for-service work, quality control testing, product release, diagnostic use, or regulatory studies - please complete the Intended Use Form so we can prepare a suitable agreement tailored to your project.Please note: The MTA applies only to certain cell lines. If this notice and the MTA document appear on a product page, the agreement is applicable. For cell lines not covered by the MTA, no reference to the agreement will be shown. The MTA is not valid for customers in the Americas, China, or Taiwan. Please contact our U.S. entity to receive the appropriate agreement. Required Product 1: 820300a Required Product 3: 860015.0 Required Product 4: 830100.0