Even if I originally try to seed the wells of a plate with what should be enough cells to completely cover the bottom of the well confluently, when I check them 48 hours later only a small percentage actually seems to have differentiated and adhered. Has my PMA just gone off, or is there a specific brand of PMA that you would recommend? Are there more effective ways to differentiate THP-1s into macrophages? Thank you for any input!
Hello Emily C. Marino
May be the PMA concentration you are using is high. The culture conditions of THP-1 such as concentration of PMA and time is extremely important before starting differentiation studies using THP-1 cells. The phenotype of the cells following PMA stimulation may vary accordingly. Please refer to the two research articles attached for more information.
Article Effect of Culture Conditions on the Phenotype of THP-1 Monoc...
Article Optimized THP-1 differentiation is required for the detectio...
I hope this helps.
Best Wishes.
Hi,
Here in our lab, we differentiate THP-1 with PMA from Sigma (P1585). I had the same problem twice. I suppose my problem was just my PMA that were gone, because I tried again with a brand new and cells differentiated well. But the second time it happened I was using also a brand new and cells did not differentiate. Once again, I tried with a brand new and it worked.
Maybe you are having the same problem. We have been working with this for so long and it happend just twice, that s why we never tried another brand.
Hope this helps!
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