Hello, everybody,
I need to treat THP-1 cells after differentiating them with PMA. I have read many articles and there is not much consensus on the conditions (seeding density, concentration of PMA, exposure time and resting phase); but well, I understand that everyone needs to optimize it. Anyway, what worries me is that almost all authors look at the expression CD14. I understand this monocyte marker should lessen its expression after PMA, but some authors see an increase and others a decrease and all interpret it as a sign of macrophage differentiation. This is confusing. In addition, the expression of CD14 in THP-1 before treatment is also very variable.
In my hands, THP1 do not express CD14 and when treated with PMA (10, 50 and 100 ng/ml) the percentage of expression increases, being greater at 10 ng/ml. In addition, cell viability appears to be declining at high concentrations of PMA.
I know that some questions have been asked about this previously, but I would like to know your opinion about whether CD14 is an appropriate marker for these cells and if you know others more robust and reliable.
Thank you very much.
Elena
I recommend to read this article before setting your experimental conditions with THP-1 cells.
http://onlinelibrary.wiley.com/doi/10.1111/aji.12129/full
Hope this is useful.
Dear Gabriel, I read the article you recommended. Curiously, I observe an intermediate case, untreated CD14 negative cells and increased marker after PMA. Anyway, they look for changes in CD14 after treatment, and I just have doubts about its usefulness. Authors suggest that CD14 could be a marker of culture quality. I'll think about it. Thank you for your input.
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