A common feature of inflammatory diseases is the accumulation of macrophages, derived from circulating monocytes, in the inflamed tissue. Rather than being a uniform cellular population, macrophages may be polarized toward pro-inflammatory (M1) or tissue reparative (M2) phenotypes. M2 macrophages polarized by anti-inflammatory cytokines are generally considered to have anti-inflammatory, anti-oxidative and tissue reparative properties. As a leading specialist in macrophage therapeutic development, Creative Biolabs excels at the identification of M2 macrophages. We are confident that our technology will meet clients' special needs.

Alternatively Activated M2 Macrophage

Alternatively activated macrophages (M2 macrophages) are a subpopulation of macrophages activated by Th2 cytokines (e.g., IL-4, IL-13) or anti-inflammatory signals (e.g., IL-10, TGF-β) with anti-inflammatory, pro-restorative, and immunomodulatory properties. Their core functions include inhibition of inflammatory responses, promotion of tissue remodeling, angiogenesis, and parasite defense, but may also be involved in tumor progression. M2 macrophages are further categorized into four subtypes based on stimulatory factors and functional differences:

Fig. 1 Advanced subtypes of M2 macrophage.^1,3

• M2a: Activated by IL-4/IL-13, secreting chemokines such as CCL17 and CCL18, recruiting Th2 cells and eosinophils, and promoting parasite immunity, wound healing and collagen deposition.
• M2b: Activated by immune complexes (IC) in combination with LPS or IL-1β, with immunomodulatory functions, secretes IL-10 and TNF-α, involved in allergic reactions and chronic inflammation.
• M2c: Activated by IL-10, TGF-β or glucocorticoids, suppresses immune response by phagocytosis of apoptotic cells and secretion of anti-inflammatory factors (e.g., TGF-β), promotes tissue fibrosis and tumor immune escape.
• M2d: Activated by adenosine, IL-6, or LPS, highly expresses VEGF and MMPs, promotes angiogenesis and tumor metastasis, and highly overlaps with tumor-associated macrophages (TAMs) phenotype.

M2 Macrophage Identification Service at Creative Biolabs

With industry-leading expertise and state-of-the-art equipment, Creative Biolabs has accumulated extensive experience in the identification of M2 macrophages by quantification of M2 macrophage markers, including multiple cell surface markers, transcription factors, and cytokine profiles.

Our core service process includes:

Our comprehensive services for human and mouse M2 macrophage identification include but are not limited to real-time PCR, liquid chromatography-tandem mass spectrometry (LC-MS/MS), western blot (WB), immunohistochemistry (IHC) and flow cytometry (FC). Moreover, our scientists have developed machine learning algorithms to automatedly identify M2 macrophage functional phenotypes using their cell size and morphology. After that, fluorescent microscopy will be conducted to assess the cell morphology of M2 macrophages.

Our identification services can be applied to the following fields:

• Disease research: M2 macrophages are closely related to the tumor microenvironment, chronic inflammation and tissue repair, and can be used to study the mechanism of related diseases.
• Drug development: To help assess the modulation of M2 polarization by drugs.
• Tissue engineering and regenerative medicine: To study the function of M2 macrophages in tissue repair.

Related Products

We provide M2 macrophage phenotypic identification services using advanced flow cytometry and multicolor fluorescent labeling technologies, which can accurately distinguish the characteristic markers and functional states of M2 macrophages and help researchers gain insight into macrophage behavior in various pathological states.

On this basis, we have also launched a series of products specifically designed for macrophage research. These products include customized kits and cellular products designed to provide researchers with comprehensive tools to support their research explorations in the field of macrophages. Our products undergo stringent quality control to ensure their efficiency and reliability in experiments, making them ideal for your related research.

Service Features

Full Process Coverage

One-stop service from monocyte isolation, polarization induction to phenotyping, supporting fresh/frozen cell delivery.

High Sensitivity Technology

Multi-parameter flow-through assay (can distinguish between M2 subtypes) with the ability to analyze low abundance molecules (e.g. exosomal miRNA).

Customized Solutions

Design specific polarization conditions (e.g. tumor microenvironment simulation) or integrate drug screening services (e.g. STAT6 inhibitor testing) according to research needs.

Publications

To lay the groundwork for the complexity of macrophage phenotypes in vivo, Kyle A. Jablonski et al. performed a comprehensive transcriptional characterization of mouse M0, M1, and M2 macrophages and identified genes common to or unique to both subpopulations. They validated the M1-exclusive expression patterns of CD38, G protein-coupled receptor 18 (Gpr18), and formyl peptide receptor 2 (Fpr2) by real-time PCR, whereas early growth response protein 2 (Egr2) and c-Myc were unique to M2.

Fig. 2 Expression of CD38 and Egr2 protein in M0, M1 and M2 macrophages.^2,3

They attempted to use these proteins as markers to differentiate between M1 and M2 macrophage populations by designing a flow cytometry panel to detect membrane CD38 and intracellular Egr2. The study demonstrated that CD38 and Egr2 are M1 and M2 markers, respectively, and that Egr2 is a better marker of M2 proteins than Arg1 or CD206.

Scientific Resources

Learn More Macrophage Biology

Download M1 vs. M2 Macrophages

Learn More Macrophages Markers

Learn More Isolation and Identification of Macrophages

Q & A

Q: What markers do you use to identify M2 macrophages?

A: We use a combination of markers to accurately identify M2 macrophages. These include CD206, CD163, CD11b, CD68, CD14, IL-10, TGF-β, Arginase-1 and others. We typically use flow cytometry to perform multiparametric analyses of both these surface markers and intracellular molecules to fully and accurately characterize M2 macrophages. In addition, we can adapt and optimize the combination of markers according to our clients' specific research needs.

Q: Can you detect different subtypes of M2 macrophages?

A: Yes, we are able to detect different subtypes of M2 macrophages, which can be further categorized into M2a, M2b, M2c, and M2d subtypes based on their activation stimulus and function. Our service can distinguish between these subtypes. We use a combination of multiparametric flow cytometry and functional analysis to differentiate these subtypes. Depending on the client's research needs, we can design specific combinations of markers and functional tests to precisely identify the M2 subtype of interest.

Q: Can you provide M2 macrophage induction and culture services?

A: Yes, we offer comprehensive M2 macrophage induction and culture services. Our services include primary macrophage isolation, M2 macrophage polarization, long-term culture, quality control, functional validation and customized services.

Q: Does your service include functional analysis of M2 macrophages?

A: Yes, our M2 macrophage phenotyping service includes not only surface marker analysis but also comprehensive functional analysis. We offer the following functional tests.

• Cytokine secretion profiling
• Phagocytosis test
• Enzyme activity assay
• Gene expression analysis
• Metabolic profiling
• Chemotaxis test

These functional analyses can be combined with surface marker assays to provide a comprehensive picture of M2 macrophage phenotype and function. We can customize the most appropriate combination of functional assays to meet your specific research goals.

References

1. Mohapatra, Swati, et al. "Non-coding RNAs regulation of macrophage polarization in cancer." Molecular cancer 20 (2021): 1-15. https://doi.org/10.1186/s12943-021-01313-x
2. Jablonski, Kyle A., et al. "Novel markers to delineate murine M1 and M2 macrophages." PloS one 10.12 (2015): e0145342. https://doi.org/10.1371/journal.pone.0145342
3. Under Open Access license CC BY 4.0, without modification.