RAW 264.7 Cells

Categories: Mouse cell lines Description: RAW 264.7 cells are a widely used murine macrophage cell line derived from the ascites of a male mouse with a tumor induced by the Abelson murine leukemia virus and are commonly used in immunological and infectious disease research. As an immortalized cell line, RAW264.7 cells are a key model system for studying macrophage biology, including immune responses to pathogens, signal transduction, and gene expression. RAW264.7 cells are particularly valuable for their ability to differentiate into macrophage-like cells. These cells can be polarized into M1 macrophages, associated with inflammatory responses, or M2 macrophages, linked to tissue repair and anti-inflammatory processes. This polarization capacity, along with their ability to perform essential macrophage functions like pinocytosis and phagocytosis, underscores their relevance in studying macrophage biology and the complex interplay between immune responses and pathogens. RAW 264.7 cells are instrumental in studying the immune system's interactions with various factors, including pathogens and bone biology. RAW264.7 cells can be induced to differentiate into osteoclast-like cells under certain conditions, such as exposure to RANKL (Receptor Activator of Nuclear Factor kappaB Ligand), making them a model for studying certain aspects of osteoclast biology and bone resorption. The RAW264.7 cell line's response to various stimuli, including the induction of pyroptosis, an inflammatory cell death process triggered by factors such as LPS (lipopolysaccharide), is instrumental in dissecting the pathways leading to inflammatory cytokine production. The impact of environmental conditions, such as extracellular glucose levels on cell function and phenotype, offers insights into cellular metabolism and the potential downregulation of inflammatory responses. RAW264.7 cells, with their origins in murine leukemia and their extensive use in immunological research, serve as a crucial tool in advancing our understanding of macrophage biology, immune system-pathogen dynamics, osteoimmunology, and inflammatory responses, highlighting their indispensable role in both basic and applied biomedical research. Organism: Mouse Tissue: Ascites Disease: Leukemia Synonyms: RAW264, RAW2647, RAW264.7, RAW-264.7, Raw 264.7, Raw264.7 Breed: BALB/c Age: Adult Gender: Male Cell Type: Macrophage Growth Properties: Adherent Citation: RAW 264.7 (Cytion catalog number 400319) Biosafety Level: 2 Ncbi_ Taxid: 10090.0 Cellosaurus Accession: CVCL_0493 Receptors Expressed: Immunoglobulin (Fc), complement (C3) Antigen Expression: H-2d Viruses: The cell line was tested and found positive for Reverse Transcriptase (RT) activity from C-Type retroviruses in cell culture supernatant and cell extract. Ectromelia virus (mousepox) may be secreted. Products: Lysozyme Culture Medium: RPMI 1640, w: 2.0 mM stable Glutamine, w: 2.0 g/L NaHCO3 (Cytion article number 820700a) Supplements: Supplement the medium with 10% FBS Dissociation Reagent: Strongly adhesive cells, use of cell scraper Doubling Time: RAW264.7 cells exhibit a doubling time ranging from 11 to 30 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: 4 x 104 cells/cm2 Fluid Renewal: 2 to 3 times per week 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. Disclaimer: Our cells are provided for in vitro laboratory research purposes exclusively and are not intended for clinical or diagnostic use, nor are they to be administered to humans or used for veterinary purposes. Users must adhere to all applicable guidelines and regulations for the handling and use of these cells in a research setting. 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. Certificate Of Analysis: The certificate of analysis can be requested on the website or via email at info@cytion.com. Please indicate the lot number of your product in the email. 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: 820700a Required Product 3: 860015.0 Required Product 4: 830100.0