RAW264.7 Cell Line Profile
Quick Facts
| Feature | Information |
|---|---|
| Cell Line | RAW264.7 |
| Species | Mouse (Mus musculus) |
| Tissue of Origin | Abelson Murine Leukemia Virus-Induced Tumor |
| Cell Type | Macrophage-like |
| Growth Mode | Adherent |
| Morphology | Round to irregular, macrophage-like |
| Recommended Medium | DMEM + 10% FBS |
| Incubation Conditions | 37°C, 5% CO₂, ≥95% humidity |
| Typical Doubling Time | Approximately 12-20 hours |
| Major Applications | Inflammation, Macrophage Biology, Phagocytosis, Osteoclastogenesis, Host-Pathogen Interactions |
Overview
RAW264.7 is one of the most widely used murine macrophage cell lines in immunology research. Unlike THP-1 and U937 cells, RAW264.7 cells naturally exhibit macrophage-like properties and do not require differentiation before use. Their rapid growth, strong inflammatory responses, and high phagocytic activity have made them a standard model for studying innate immunity, inflammatory signaling, and macrophage function.
Origin and History
- Derived from a tumor induced by Abelson Murine Leukemia Virus in mice.
- Established as a continuously growing macrophage-like cell line.
- Extensively used in immunology, inflammation, biomaterials, and bone biology research.
Growth Characteristics
| Characteristic | Description |
|---|---|
| Growth Mode | Adherent |
| Morphology | Round to irregular, macrophage-like |
| Growth Pattern | Forms an adherent monolayer |
| Doubling Time | Approximately 12-20 hours |
Culture Notes
RAW264.7 cells adhere firmly to culture vessels but are sensitive to harsh enzymatic detachment. Many laboratories use gentle scraping or minimal trypsin exposure to preserve cell viability and macrophage function.
Biological Characteristics
Inflammatory Activation
RAW264.7 cells respond robustly to inflammatory stimuli, particularly Lipopolysaccharide (LPS), which activates TLR4 signaling and induces strong inflammatory responses.
Following activation, cells typically:
- Increase cytokine production
- Produce nitric oxide (NO)
- Undergo morphological changes
- Activate NF-κB and MAPK signaling pathways
Nitric Oxide Production
A defining feature of RAW264.7 cells is their ability to express inducible Nitric Oxide Synthase (iNOS) and produce high levels of nitric oxide following stimulation. This makes them one of the most widely used models for studying nitric oxide biology and inflammatory signaling.
Phagocytosis
RAW264.7 cells possess strong phagocytic activity and are frequently used to investigate:
- Particle uptake
- Bacterial clearance
- Nanoparticle interactions
- Innate immune recognition
Osteoclast Differentiation
Unlike many immune cell lines, RAW264.7 cells can differentiate into osteoclast-like cells following stimulation with RANKL.
This property makes them valuable for studying:
- Bone remodeling
- Osteoporosis
- Osteoclast biology
- Arthritis
Culture Conditions
Recommended Medium
- DMEM
- 10% Fetal Bovine Serum (FBS)
Incubation Conditions
| Parameter | Value |
|---|---|
| Temperature | 37°C |
| CO₂ | 5% |
| Relative Humidity | ≥95% |
Passaging
RAW264.7 cells should be passaged before reaching complete confluency. Gentle handling is recommended to preserve macrophage phenotype and viability.
Major Applications
RAW264.7 cells are widely used for:
- Macrophage biology
- Inflammation research
- Cytokine signaling
- Nitric oxide studies
- Phagocytosis assays
- Host-pathogen interactions
- Osteoclast differentiation
- Biomaterials research
- Drug discovery
- Cancer immunology
Advantages
- Naturally macrophage-like without differentiation
- Strong and reproducible inflammatory responses
- High phagocytic activity
- Rapid growth and easy maintenance
- Extensive experimental literature
Limitations
- Mouse-derived cell line with species-specific differences
- Tumor-derived rather than primary macrophages
- Certain signaling pathways differ from primary immune cells
- Simplified model lacking tissue microenvironment
Authentication and Quality Control
Cell Line Authentication
Routine authentication through species verification and morphological assessment is recommended. Functional activation assays may also be used to confirm macrophage phenotype.
Contamination Monitoring
Regular screening should include:
- Mycoplasma contamination
- Bacterial contamination
- Fungal contamination
Routine quality control is essential for reliable and reproducible experimental results.
Cryopreservation
RAW264.7 cells generally recover well following standard cryopreservation and thawing procedures. Maintaining low-passage master stocks is recommended to preserve consistent inflammatory responses.
References
- Freshney RI. Culture of Animal Cells: A Manual of Basic Technique and Specialized Applications.
- ATCC Cell Line Database.
- Cellosaurus Cell Line Knowledgebase.
- ECACC Cell Line Collection.