The alternatively activated, anti-inflammatory M2 macrophages can be separated into at least three subgroups (M2a, M2b, and M2c), which have different functions, including regulation of immunity, maintenance of tolerance and wound healing. Creative Biolabs is well equipped and versed in M2a, M2b, and M2c macrophage polarization assay. Our assay is also designed to evaluate the cell surface receptor expression, cytokine profiles, scavenging functions, and ability to activate or suppress T-cell proliferation.

M2 Macrophage Polarization

M2 macrophage polarization is a functional phenotypic switching process that occurs in macrophages in response to specific microenvironmental stimuli, and is mainly characterized by anti-inflammatory, pro-repair and immunomodulatory properties. Unlike pro-inflammatory M1-type macrophages, the M2 type is usually induced by Th2-type cytokines (e.g., IL-4, IL-13, IL-10) or immune complexes.

Fig. 1 M2 macrophage polarization pathways.^1,3

Based on inducing factors and functional differences, M2 types are further categorized:

• M2a: Induced by IL-4/IL-13, high expression of CD206, Arg1, promoting tissue repair and fibrosis.
• M2b: Activated by immune complexes and TLR ligands, secretes IL-10 but low expression of IL-12, involved in immune regulation.
• M2c: Induced by IL-10 or glucocorticoids, high expression of CD163, removes apoptotic cells and inhibits inflammation.
• M2d: Tumor-associated macrophage, promotes angiogenesis and tumor immune escape.

Key factors that trigger M2 polarization include:

• Cytokines: IL-4, IL-13, IL-10, and TGF-β are the main inducers.
• Metabolites: Butyrate, PGE2 promote M2 polarization by enhancing OXPHOS.
• Drugs and physical stimuli: Statins, nanostructured titanium surfaces can induce M2 phenotype by modulating cell signaling or morphology.
• Transcription factors: STAT6, PPARγ, and C/EBPβ are coregulatory molecules.

M2 Macrophage Polarization Assay at Creative Biolabs

M2 macrophage polarization assay service is designed to accurately assess the polarization status of macrophages under specific stimuli through multi-dimensional technical means, providing key data support for studies on immune regulation, tumor microenvironment, tissue repair and other research.

Our service aims to help researchers gain a deeper understanding of the polarization state of M2 macrophages and their roles in different biological processes, covering the complete process from cell polarization induction, phenotypic identification to functional analysis.

Table 1 Service process of M2 macrophage polarization assay

Through our technical system and service design, this assay service is able to provide research users with high-precision and high-reliability M2 macrophage polarization data, and help immune microenvironment mechanism research and translational medicine exploration, for example:

• Tumor immune research: Detecting the M2/TAM ratio of tumor tissues and assessing the efficacy of PD-1/CTLA-4 inhibitors.
• Fibrotic diseases: Analyzing the anti-fibrotic function of M2c macrophages in lung/liver fibrosis models.
• Regenerative medicine: Assessing the pro-angiogenic role of M2a macrophages in bone repair or wound healing.

Related Products

M2 macrophages play a key role in maintaining tissue homeostasis, promoting repair and regulating immune responses. Therefore, accurate detection and analysis of M2 macrophage polarization status is important for understanding the mechanisms of numerous diseases.

As our services continue to evolve, we are also pleased to introduce a number of innovative products, including monocytes, macrophages and various assay kits. We are committed to providing researchers with comprehensive and systematic solutions.

Below are some of our popular products. You can click to view the details.

Service Features

Subtype Segmentation

Our services support M2a-M2d subtype-specific assays to meet the needs of tumor immunology, fibrosis, and other segmented areas.

Technology Integration

Our multi-methods integration, combined with flow, single-cell sequencing and metabolic analysis, provides multi-dimensional data cross-validation.

Disease Model Adaptation

Optimize polarization scheme and detection indexes for tumor microenvironment, chronic inflammation, tissue repair and other scenarios.

High-throughput Platform

We are able to handle multiple samples, which is suitable for large-scale research projects.

Customized Solutions

We provide personalized assay protocols and data analysis according to customers' specific research purposes.

Fast Delivery

Rapid data analysis and result reporting help researchers obtain timely experimental results.

Publications

Zhao, Chongru et al. investigated the effect of Cancer-associated adipocytes (CAA)-derived IL-6 on macrophage polarization in promoting breast cancer (BC) progression. They used functional assays, qRT-PCR, Western blot and ELISA to explore the effects of CAA-derived IL-6 on macrophage polarization and PD-L1 expression.

Fig. 2 CAA-derived IL-6 induce M2 macrophage polarization by activating STAT3.^2,3

The results revealed that CAA could secrete abundant IL-6, thereby inducing M2 macrophage polarization through activation of STAT3. In addition, CAA could upregulate PD-L1 expression in macrophages.

Scientific Resources

Learn More Macrophage Biology

Download Macrophage Polarization

Learn More Different Approaches to Regulating Polarization of Tumor-Associated Macrophages

Learn More Isolation and Identification of Macrophages

Q & A

Q: What are the sample types and delivery requirements? Will the results be affected if there is insufficient cell activity?

A: We recommend the use of primary macrophages or the RAW264.7 cell line with ≥85% viability for samples. Single cell suspension preparation of fresh tissue is subject to collagenase digestion and DNase I treatment. A viability of <80% may result in down-regulation of M2 marker expression, and it is recommended that a cell activity assay be provided. If the viability is insufficient, we can provide sample pretreatment services.

Q: What is the delivery time for test results and will a detailed report be provided?

A: The exact timing of the test depends on the number of samples and the complexity of the experiment. Upon delivery of the results, a detailed lab report will be provided, including quantitative analysis of the assay results, graphs and data interpretation. The report will provide a comprehensive description of the polarization status of M2 type macrophages, which will be easy for you to refer to and further analyze in subsequent studies. If you have any special requirements or wish to obtain additional analysis, we will be happy to help you meet them.

Q: What types of samples can I provide for testing?

A: We can accept many types of samples, including PBMC, tumor tissue sections, primary cell cultures and other samples. Each sample type may require different handling and storage conditions. We encourage our clients to contact us prior to submitting their samples to ensure that we can provide the best possible service and advice.

Q: How do I get started with your testing services? What is the whole process?

A: Getting started with our inspection services is very simple. First, you can obtain a service description and quote by visiting our website or contacting customer service directly. Next, we will discuss with you the specific needs of your experiment, including sample type, assay and schedule. Once agreed upon, you can submit your samples according to the sample handling guidelines we provide. We will begin testing within a predetermined time frame after receiving the sample and provide a report of the results upon completion. The entire process is transparent so that you are kept informed of progress.

References

1. Martins, Rubens Andrade, et al. "Regenerative Inflammation: The Mechanism Explained from the Perspective of Buffy-Coat Protagonism and Macrophage Polarization." International Journal of Molecular Sciences 25.20 (2024): 11329. https://doi.org/10.3390/ijms252011329
2. Zhao, Chongru, et al. "CAA-derived IL-6 induced M2 macrophage polarization by activating STAT3." BMC cancer 23.1 (2023): 392. https://doi.org/10.1186/s12885-023-10826-1
3. Under Open Access license CC BY 4.0, without modification.