RAW 264.7 Cell Line in Macrophage and Immunology Research

Let's start with RAW 264.7 cells, a powerful tool in the study of mouse macrophages. The RAW264.7 monocyte/macrophage-like cell line is commonly used to investigate how host cells react to microorganisms and their byproducts. RAW 264.7 cells also are 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.

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.

Cellular Functions and Interactions

RAW 264.7 cells, as antigen-presenting cells, play a pivotal role in the immune system by processing and presenting antigens to T cells, thereby initiating immune responses. This cell interaction is crucial for understanding how the immune system recognizes and responds to pathogens. The cell viability and cellular actions of RAW 264.7 cells under different conditions provide key insights into macrophage functions, such as phagocytosis and cytokine production. These cells also serve as a model for studying cellular aging and intracellular age levels, which can impact immune responses.

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].

Cell culture information on 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!

Pioneer your research with our RAW 264.7 cells

Advantages and disadvantages of RAW 264.7 cells

Advantages

• 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.

Disadvantages

• 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.

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].

Osteoclastogenesis model

In the context of osteoclastogenesis, RAW 264.7 cells have been used to explore the differentiation of macrophages into osteoclast-like cells, albeit with certain limitations due to their monocyte lineage. The study of osteoclast behavior, osteoclast differentiation, and related processes in RAW 264.7 cells contributes to our understanding of bone resorption and the pathogenesis of osteoporosis. Chemical tools and fluorescence-based imaging techniques, including optical imaging, are extensively applied to visualize intracellular fluorescent age formation and investigate heme metabolism within these cells.

Molecular Insights and Osteoclastogenesis

While primarily a macrophage model, RAW 264.7 cells have been employed in vitro osteoclastogenesis studies to gain insights into osteoclastogenic differentiation capacity and gene expression. Although not a direct model for osteoclasts, the myeloid cell line can be treated with various factors to induce osteoclast-like features, aiding in the understanding of osteoclast lineage and osteoclast precursor cells. This application, however, should be approached with caution, considering the cell line's origin and inherent characteristics.

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].

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.

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.

• Transfecting RAW 264.7 cells: This website will provide a protocol for transfecting RAW 264.7 macrophages using a lipofectamine reagent.
• Transfection protocol: This published article describes a detailed protocol for the transfection of RAW 264.7 cells.
• Efficient Transfection method for RAW 264.7 cells: This document describes another practical transfection approach using RAW 264.7 cells.

Videos related to RAW 264.7 cell line

• Culturing RAW 264.7 cells: This video explains RAW 264.7 cells passaging protocol.
• Transfection method: This video is about transfecting RAW 264.7 cells with a reporter gene.

Research publications using RAW 264.7 cells

Many research publications are available on mouse macrophage cell line RAW 264.7. 

• Polysaccharides from Polygonatum sibiricum Delar. ex Redoute induce an immune response in the RAW264. 7 cell line via an NF-κB/MAPK pathway: This study proposed that polysaccharides obtained from the Polygonatum sibiricum plant can trigger an immune response in the mouse macrophage cell line RAW 264.7 via regulating NF-κB/MAPK signaling cascade.
• Immunomodulatory effects of Tinospora crispa extract and its major compounds on the immune functions of RAW 264.7 macrophages: This article published in the International Immunopharmacology journal describes thatTinospora crispa plant extract and its major constituents have an immunomodulatory and immune-stimulatory effect on the RAW 264.7 macrophage cell line.
• Heat-killed lactic acid bacteria inhibit nitric oxide production via inducible nitric oxide synthase and cyclooxygenase-2 in RAW 264.7 cells: This study proposed that heat-killed lactic acid bacteria can potentially be used in probiotic product synthesis because they possess immune-modulatory and antioxidant properties. Researchers used RAW 264.7 cells to study these biological activities.
• Effect of herbal formulation on immune response enhancement in RAW 264.7 macrophages: This study used RAW 264.7 cells to study the immune-stimulatory effect of an herbal formulation named KM1608. The study suggested KM1608 as a potential immune response enhancement agent.
• A screening of plants used in Colombian traditional medicine revealed the anti-inflammatory potential of Physalis angulata calyces: This paper published in the Saudi Journal of Biological Sciences investigated the anti-inflammatory potential of 10 commonly used traditional Colombian plants using LPS-stimulated RAW 264.7 cells.

References

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.