RAW264.7 cells are a widely used laboratory model in biological research. These mouse-derived immune cells, specifically macrophages, are employed to understand various cellular processes and diseases.
Understanding RAW 264.7 Cells
RAW 264.7 cells originated from a male BALB/c mouse, specifically from a tumor induced by the Abelson murine leukemia virus (A-MuLV) in the 1970s. This viral transformation immortalized the cells, allowing them to proliferate indefinitely in a laboratory. They are a macrophage-like cell line.
These cells primarily grow adherently, attaching to culture dish surfaces. Their morphology varies, appearing round or spindle-shaped. RAW 264.7 cells produce nitric oxide (NO), perform phagocytosis, and respond to immune stimulants.
They are cultured in Dulbecco’s Modified Eagle’s Medium (DMEM) supplemented with 10% fetal bovine serum (FBS) and 1% penicillin-streptomycin, maintained at 37°C with 5% CO2. This provides a stable and reproducible system for studying macrophage behavior.
Unique Features for Scientific Study
RAW 264.7 cells are valued for their ease of culture, robustness, and consistent growth, which leads to reproducible experimental outcomes. They are subcultured by scraping when 70-80% confluent. Their average doubling time is approximately 15 hours, ensuring a steady cell supply.
A key feature is their ability to be stimulated by various substances, mimicking immune responses. For instance, bacterial components like lipopolysaccharide (LPS) induce an inflammatory response, increasing nitric oxide production and phagocytosis. They can also be polarized into different macrophage phenotypes, such as M1 (pro-inflammatory) or M2 (anti-inflammatory), depending on the stimuli.
Their immortalized nature provides a continuous supply of identical cells, a significant advantage over primary cells that vary between isolations. This consistency reduces experimental variability and costs, making them an efficient in vitro model. They express macrophage-specific surface markers like F4/80 and CD14, confirming their macrophage-like phenotype.
How RAW 264.7 Cells Aid Research
RAW 264.7 cells are extensively used in immunology research, especially for understanding inflammatory responses and immune signaling pathways. When stimulated with bacterial endotoxins like LPS, these cells produce pro-inflammatory cytokines such as interleukin-6 (IL-6), interleukin-1β (IL-1β), and tumor necrosis factor-alpha (TNF-α), and prostaglandin E2 (PGE2). This allows scientists to investigate how immune cells react to triggers and explore inflammation mechanisms. For example, studies use them to examine how compounds suppress inflammatory mediators by inhibiting pathways like NF-κB.
These cells also aid drug discovery and screening, particularly for identifying anti-inflammatory agents or immunomodulators. Researchers treat LPS-stimulated RAW 264.7 cells with new compounds to observe their ability to reduce inflammation at the cellular level, by measuring decreased NO or cytokine production. This helps find new therapeutic candidates for inflammatory diseases, as demonstrated in studies screening natural product extracts.
RAW 264.7 cells are also used in toxicology to study cellular responses to toxins, environmental pollutants, or nanoparticles. For example, they assess the cytotoxicity and inflammatory effects of silica or zinc oxide nanoparticles, demonstrating sensitivity and particle accumulation capacity. This aids understanding potential adverse health outcomes from material exposure.
Their utility extends to studying pathogen-host interactions, such as how immune cells recognize and respond to bacteria or viruses. By infecting RAW 264.7 cells with pathogens like Mycobacterium tuberculosis or mouse hepatitis virus, researchers can analyze the cellular immune response, including gene expression changes, providing insights into host defense mechanisms.