I was wondering how I could induce the M2 phenotype and specifically CD206 expression in RAW264.7 cells? I read that treating RAW264.7 cells with IL-4 did not lead to expression of CD206. If IL-4 does not lead to expression of CD206, then I was wondering if there is another way I can induce expression of CD206 in the RAW264.7 cell line. Thank you in advance!
Hi,
The M2 phenotype, also known as the alternative activation phenotype, is characterized by the expression of markers such as CD206. The RAW264.7 cell line is a murine macrophage cell line, and it can be induced to adopt an M2 phenotype by treatment with certain cytokines and other molecules.
One way to induce the M2 phenotype in RAW264.7 cells is to treat them with interleukin-13 (IL-13), which has been shown to upregulate the expression of CD206 in these cells. Other molecules that can be used to induce M2 polarization in RAW264.7 cells include IL-4, transforming growth factor-beta (TGF-β), and insulin-like growth factor-1 (IGF-1). However, as you mentioned, treatment with IL-4 alone may not lead to expression of CD206, indicating IL-4 might be not optimal choice.
Some researcher also used combinations of various cytokines and growth factors along with activators like LPS to achieve the desired phenotype. It's also important to note that different cell lines and culturing conditions could affect the outcome of the polarization.
It's also recommended to check the literature and/or to reach out to scientists working on similar cell lines for more specific protocols for your particular case.
Regards.
Hi, generally RAW cells are M1 biased and can easily be polarized to M1. Activating them to M2 phenotype is a bit tough in my experience so we generally work with Bone marrow derived macrophages for such polarisation experiments.
However, you can use a cocktail of IL-4 (10ng/ml) + Il-10 (10ng/ml) or TGF-b (10ng/ml) to activate RAW towards M2. You can give M2 stimulus for 12-24 hrs and then can check for the marker.
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