Hi, I'm new to this cell line, but how actually they look like before and after PMA-induction? I saw tiny 'creatures' in the culture media, not sure they are swimming in or not (moving in a confined space), but can see some attaching to membrane, some located intracellularly, under 40X magnification. Are those things bacteria contaminants or other stuff? I have check the media & FBS turbidity, and spread them as well as cell supernatant on NA plate but all show negative (no turbid & no cfu over days).
Hi Zhi Kho,
I don´t see contaminations, but the cells looks not fine. I see cells to die off and cells in apoptose I suppose.
Jochem
There is no contamination either in control or PMA treated cells. The change in morphology of cells is because of PMA treatment. As Zhi Kho has mentioned, there are dead cells in the treated cells. You may have to lower the concentration of PMA or reduce the treatment time to avoid cell death.
Agree with the above. Your "activated" cells are dead. The "little" particles are cell debris.
By the way, your control THP-1, what did you use for differentiation?
Thanks for the recommendation and reply! Shen-An Hwang I use 25nM PMA 48h, 24h resting. Btw does the monocyte looks healthy? i think they might be apoptose also (blebbing observed).
Yeah.. your monocyte cultures (the bottom picture), the cells do not look healthy either.
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