Hi! I am trying to differentiate human monocyte to macrophages using M-CSF but I am having some problems! There are lots of papers but much of them indicates different concentration of M-CSF.
Which is the best concentration? I Want to polarize the macrophages to M1 and M2 after differentiation...When I did the polarization (48 hs LPS 100 ng/ml IFNy 20 ng/ml ; IL4/IL13 20 ng/ml respectively) after 7 day with 50 ng/ml M-CSF all of them are M2 (up regulation of CD206 by FACs)
Can you help me?
Thaks!!!
Do not use M-CSF. Use conditioned media from LM-929 cells (which contains M-CSF too). Adding this media to monocytes will turn into macrophages. This is a well-recognized method and frequently used.
Hello
I use M-CSF on monocytes (CD14+ cells within PBMC) for 6 days. The last 3 days I add LPS/IFNg or IL4 or IL10 to produce M1, M2a or M2c respectively. The medium is also important: I use X-vivo10 (Giraud FR PlosOne 2012).
Kind regards.
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