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RAW-264.7 Cell Line - Contribution of RAW-264.7 cells to Macrophage and Immunology Research

Let's start with RAW 264.7 cells, a powerful tool in the study of mouse macrophages. This incredible monocyte/macrophage-like cell line is commonly used to investigate how host cells react to microorganisms and their byproducts. But that's not all – RAW 264.7 is also an excellent tool for evaluating the potential therapeutic benefits of natural products and their bioactivity.

But before you start experimenting, it's essential to understand the basics of this remarkable cell line. Where do RAW 264.7 cells come from, and how do you culture them? These critical questions ensure you get the most out of your research. So let's discover everything you need to know about RAW 264.7 cells.

  1. What is the RAW 264.7 cell line?
  2. Cell culture information of RAW 264.7
  3. Advantages and disadvantages of RAW 264.7 cells
  4. Applications of RAW 264.7 cells
  5. RAW 264.7 cells: Resources, Protocols, Videos & More
  6. Research publications using RAW 264.7 cells

1.         What is the RAW 264.7 cell line?

The RAW 264.7 cell line is a macrophage-like cell line that was initially derived from the ascites of a tumor induced by Abelson Leukemia Virus (A-MuLV) in a male BALB/c mouse. This cell line is considered a suitable in-vitro model for macrophages due to their ability to perform phagocytosis and pinocytosis, essential functions of macrophages in the body [1]. RAW 264.7 cells are also an immortalized cancer cell line, frequently used in research due to their ability to regulate bone remodeling, although they are not osteoclasts [2].

The RAW 264.7 cell line exhibits semi-adherent growth characteristics, with spindle or cuboidal-shaped adherent cells and spherical or round-shaped floating viable cells. The cell size of RAW 264.7 cells typically ranges from 10 to 20 µm in diameter.

Are RAW264.7 cells M1 or M2 macrophages?

Macrophages can be classified into three main types: M0, M1, and M2. M0 macrophages are nonactivated, while M1 macrophages are pro-inflammatory and M2 macrophages are anti-inflammatory [3]. RAW 264.7 M0 cells can polarize into either M1 or M2-type macrophages [4].

2.        Cell culture information of RAW 264.7

Before you start your experiments, you must understand some basic information about this powerful cell line. Do you know the doubling time of RAW 264.7 cells? What's the seeding density for these cells, and are they adherent? Moreover, what are the optimal growth conditions for RAW 264.7? Keep reading to uncover all the answers you need to work with this incredible cell line!

Cell Culture Information

RAW 264.7 Cell Line

Population doubling time

11 to 30 hours

Adherent or in suspension

Mostly adherent with some suspension cell populations

Seeding density

4 x 10^4 cells/cm^2

Recommended growth medium

RPMI 1640 supplemented with 10% fetal bovine serum and L-glutamine (2.5 mM)

Optimal growth conditions

Humidified incubator with 5% CO2 and 37°C temperature

Media replacement

2 to 3 times a week


Vapor phase of liquid nitrogen

Freezing medium

CM-1 or CM-ACF

Freezing process

Slow freezing

Thawing process

Rapid agitation in 37°C water bath

Biosafety level

BSL-2 laboratory recommended


3.        Advantages and disadvantages of RAW 264.7 cells


  • Easy culturing: The RAW 264.7 cell line is easy to grow and maintain in the laboratory without any complicated requirements.
  • Well-characterized: RAW 264.7 is a well-characterized cell line, which means it has stable phenotypic and functional features.
  • In vitro model of macrophages: As RAW 264.7 cells are macrophages, they exhibit essential macrophage-like functions such as phagocytosis and pinocytosis. Thus, they are often used as an in vitro model of macrophages.
  • Differentiation to osteoclasts: RAW 264.7 cells can be differentiated into osteoclasts using specific molecular factors. These differentiated cells are capable of performing bone remodeling, similar to osteoclasts.


  • Poor representation of primary macrophages: As a transformed cell line, RAW 264.7 may not accurately represent the features of primary macrophages and may have some differences in gene expression, phenotype, and function.
  • Limitations in drug screening: RAW 264.7 cells may not be suitable for all types of drug screening assays, and their responses to some drugs may differ from primary macrophages.
  • Strong cell adhesion: RAW 264.7 cells strongly adhere to the surface of the culture flask and can sometimes be tricky to detach.

4.       Applications of RAW 264.7 cells

Study of cellular responses

Due to their similarity to macrophages, RAW 264.7 cells are frequently employed as an in vitro model to investigate cellular responses to microbes and other biological products. For instance, a study utilized RAW 264.7 cells to evaluate heat-killed lactic acid bacteria's immunomodulatory and antioxidant activity [5].

In another research conducted in 2019, the immunological activity of polysaccharides derived from Polygonatum sibiricum plant was examined on macrophage-like RAW 264.7 cells. The findings of this study demonstrated that the polysaccharides induced an immune response by activating the NF-κB/MAPK cell signaling pathways [6].

Screening of natural products

The RAW 264.7 cell line is widely utilized to screen natural products, including plant-based compounds, for their potential bioactivity and effects. For instance, a study conducted by Korean researchers employed RAW 264.7 mouse macrophages to explore the immune-stimulating effect of a herbal formulation [7].

5.       RAW 264.7 cells: Resources, Protocols, Videos & More

RAW 264.7 is an extensively used macrophage cell line. Therefore, many online resources are available for learning about its culture and maintenance.

Cell culture protocols

Here are a few online line resources explaining RAW 264.7 cell culturing.

  • Culturing RAW 264.7 cells
  • : This link provides information about RAW 264.7 cells, including their growth medium and subculturing protocol.
  • RAW 264.7 cell line
  • : This document contains all information essential for the propagation and culture of RAW 264.7 cells.

Transfection protocols

RAW 264.7 cells are often used in transfection studies. The following resources explain different transfection methods used for the RAW 264.7 cell line.

Videos related to RAW 264.7 cell line

We hope this article has provided you with fundamental and advanced knowledge about the RAW 264.7 cell line. If you are considering utilizing these macrophage cells in your research and wish to purchase them, kindly consider ordering from us.

6.       Research publications using RAW 264.7 cells

Many research publications are available on mouse macrophage cell line RAW 264.7. Here, this section will only include some notable examples.


  1. Taciak, B., et al., Evaluation of phenotypic and functional stability of RAW 264.7 cell line through serial passages. PloS one, 2018. 13(6): p. e0198943.
  2. Wang, S., et al., Inflammatory macrophages interrupt osteocyte maturation and mineralization via regulating the Notch signaling pathway. Molecular Medicine, 2022. 28(1): p. 102.
  3. Orekhov, A.N., et al., Monocyte differentiation and macrophage polarization. Vessel Plus, 2019. 3: p. 10.
  4. Khabipov, A., et al., RAW 264.7 macrophage polarization by pancreatic cancer cells–a model for studying tumour-promoting macrophages. Anticancer Research, 2019. 39(6): p. 2871-2882.
  5. Kang, C.-H., et al., Heat-killed lactic acid bacteria inhibit nitric oxide production via inducible nitric oxide synthase and cyclooxygenase-2 in RAW 264.7 cells. Probiotics and Antimicrobial Proteins, 2021. 13(6): p. 1530-1538.
  6. Zhang, J., et al., Polysaccharides from Polygonatum sibiricum Delar. ex Redoute induce an immune response in the RAW264. 7 cell line via an NF-κB/MAPK pathway. RSC advances, 2019. 9(31): p. 17988-17994.
  7. Trinh, T.A., et al., effect of herbal formulation on immune response enhancement in RAW 264.7 macrophages. Biomolecules, 2020. 10(3): p. 424.


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