Hello every one, I currently working on THP-1 cell line, I want to convert them to macrophage like cells with adding PMA. Does any one know is necessary to prove that they are macrophages with flowcytometry? If is that necessary which marker I should use?
Though it was shown by many researchers in the past that THP-1 cells can be differentiated to macrophage-like cells upon PMA stimulation, it is always advisable to reconfirm and show the same from your end. U can show the same either with FACS, qPCR, IF or WB. Simple way is to show the changes in morphology by imaging.
If you want to go with flowcytometry, staining for markers like CD36, CD54, CD11b, HLA-DR or CD14 will be helpful. Note that PMA treatment has been reported to cleave CD14, you can get CD14 expression using vitamin D3 treatment (4-5 days) but the cells are reported to be monocyte-like rather that macrophage-like. Apparently, best markers to try would be CD11b and CD36.
Dear Molaghaee
The shape of cells will be changed (after PMA) and you can easily evaluate them by using microscope . You can do flow but it is not much necessary.
Good luck
Yes, you need to prove that the converted cells are macrophages by any method (fluorescence microscopy/Flowcytometry/WB/ PCR).
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