I plan to check the impact of exogenous cytokines in alteration of M1 or M2 phenotype using mouse macrophages. I'm not familiar with the mouse macrophage cell line. For this purpose, which cell line is suitable? I will determine phenotypes by flow (CD86, IA/IE; M1, MR; M2) and iNOS/Arginase-1 ratio by qPCR.
Thank you, Carl. Does RAW 264.7 have a neutral phenotype in terms of M1 or M2 in the conventional culture?
Some interesting data on Raw activation states. It is microglia focussed but there is data.
Thank you, Bobbi. I'm working in skin immunology. Do you think if there is any conflict to use RAW 264.7 as macrophages in skin? By the way, now I'm a fellow of NIH, USA. Last friday, I heard a wonderful lecture by Fiona Watt, King's College London.
I would recommend the IC-21 cell line. Out of 8 mouse monocyte/macrophage cell lines we screened, the IC-21 cell line responded most like an M2 cell type when challenged with inflammatory microcrystals, whereas RAW264.7 cells responded like a classical M1. IC-21s will also express arginase.
Yagnik, D., Hillier, P., Marshall, D., Krausz, T., Haskard, D.O. and Landis, R.C. “Non-inflammatory phagocytosis of monosodium urate monohydrate crystals by mouse macrophages: Implications for the control of joint inflammation in gout” Arthritis Rheum. 43 (2000), 1779-89
Please be advised that the RAW cell line and most other mouse macrophage cell lines express an XMRV like virus , meaning that they must be handled under BL2 conditions. Also, for reasons that are not clear, most macrophage lines do not express IL-12 following LPS stimulation. My lab and the lab of Jill Suttles have derived many macrophage cell lines from WT, KO and transgenic mice using the myc-raf oncogenes. However I have not looked at their M1/M2 state although they are responsive to LPS and IFN-gamma.
Can I use these cell lines for testing drugs for the anti leishmaniasis
May I know why RAW 264.7 macro phage is name as RAW 264.7? What is the history background? Thank.
We have developed mouse macrophage cell lines from many strains of mice using the raf/myc oncogene pair. The cells appear to resemble primary macrophages with one exception in that they don't make IL-12 (but neither does RAW).
The RAW 264.7 cell line is one of the best characterized cell lines representing the M1 phenotype. For the M2 phenotype, the IC-21the cell line is a good representative line(Yagnik, Landis. Arthritis Rheum. 2000 Aug;43(8):1779-89)
In our lab, the RAW 264 macrophage cells responded to fatty acid treatment and lipopolysacharides (LPS) much better than j774a cells. The RAW cells also appear to have a much better lox 1 receptor response to oxidized LDL low density lipoproteins.
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