Hello fellow researchers,
Quite recently, we differentiated HL-60 cells into PMNs using DMSO and had a fair amount of success. I usually check for the increase in surface CD11b and also control for CD14, so that I can exclude monocytes.
This time I need to develop them into Monocytes/Macrophages. Hence, I would look for CD14+ cells. But i have the following concerns:
1. How do I judge Monocyte or Macrophage stage since as far as I read Monos are CD14+ but when then turn into Macros they express more CD11b. But do they express CD11b in Monos stage?
2. In our experience, HL-60 when turned into PMNs are CD11b+/CD14 - and also CD66b - , which is strange since PMNs from fresh blood have a very good presentation of CD66b and is the classical marker for us to test for them.
So, if Macros from HL-60 will be CD14-low, CD11b-high, how do I tell them apart from PMNs?
I would appreciate any suggestions.
Thanks,
Sudip
Perhaps you could include other markers that can identify differentitated monocytes. These would include CD206 and CD163. Using FSC/SSC, you could also roughly distinguish monocytes from macrophages, since monocytes are smaller in size than macrophages. Hope this helps..
Perhaps you could include other markers that can identify differentitated monocytes. These would include CD206 and CD163. Using FSC/SSC, you could also roughly distinguish monocytes from macrophages, since monocytes are smaller in size than macrophages. Hope this helps..
That depends which kind of agent you are going to use for inducing mono/mq differentiation in HL-60 cells. Please see the attached paper (especially the figure 2 in the supplement). Note: Addition of IFN-g leads to a nice CD14-positivity and monocyte-like differentiation in HL-60 cells.
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