Hi ! MCSF set as a golden standard for differentiating human PBMC derived monocytes to macrophages (M0?) . However , in my experiments, MCSF gave macrophages more skewed towards M2 like state (with monocytes isolated from fresh PBMCs). I found M0 expression less than CD206 expression after MCSF treatment for 7 days. Has anyone also encountered this ?
It may not be very surprising since some papers (older) reported using MCSF as a M2 macrophage inducer.
I wonder how to keep macrophages restricted in complete M0 state (or atleast more than 90% in flow),since all these sub-populations are highly dynamic in nature .This would help in efficient polarization to M1/M2 by specific cytokines. Any insights would be highly appreciated .
Thanks in advance !
Hi Neha Rana ,
As you must be aware already, there are several ways to differentiate macrophages. As you mentioned M-CSF is a good way to differentiate macrophages; however, it can also be achieved with GM-CSF and even with autologous serum (or human serum).
I have certainly seen that MCSF macrophages are more M2 like while GM-CSF are more proinflammatory.
Surface markers can certainly change based on the purification method of the monocytes, culture method, serum used, etc...
If you manage to keep the macrophages, sterile, un-stress and happy they will be M0, and you can corroborate this by activating your M0 to M2 and compare the receptors.
Hi Samuel J Lara Reyna , Thanks for the response.
But even when macrophages look happy and their is a lawn (MCSF treated for 7 days), do you think around 50% CD68+ expression is enough to call them M0? What other markers can be used to justify the M0 expression or to prove that they are unpolarized?
I have also read studies where both MSCF and GMCSF are used to obtain a more homogenous M0 population before inducing M1 or M2 . Have you tried this?
Regards
Neha
The best way to corroborate if your macrophages are not activated is by activating them, and have a positive and negative control.
For instance, I normally use CD64, as a differentiation marker for M0 macrophages, and always activate them to further corroborate that they are indeed in a M0 state. I select CD64+ cells to further gate, into M1 or M2.
Then you can use CD80 and CD86 to corroborate that the macrophages are inactive/active (in the case of M1) or CD209 and CD206 for M2. Several other markers exist for macrophages and these are not the only ones.
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