Hi. Can anyone help.
I induce THP-1 cells into M1 macrophages.
Firstly, I use PMA 50ng/ml to induce THP-1 cells into M0 macrophage for 24h, the cells will attach and distributed evenly on dish surface,
I then replace the media with PMA 50ng/ml, IFN-r 20ng/ml, LPS 1ng/ml for another 18h to induce M0 into M1, but I found that many M1 cells clump together, as shown in the attachment. Can anyone tell why M1 cells clump, Is the THP-1 not in good condition, or I use a high dose of IFN-r concentration, or other reasons? thanks.
Dear Wei, Your differentiated cells look healthy. There is nothing to work. Regarding use of PMA, as per my experience, a dose of 5ng/ml for 6-8h is enough.
Thanks Harsh, you mean it is normal that M1 macrophages will clump like this. But can I avoid them clumping in the differentiation?
i would also suggest using a lower dose of PMA. I used to use 30ng/ml for THP-1 cells to convert them to M1 macrophages. if the problem persists you can change the THP-1 batch of cell.
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