I activated macrophages derived from U937 with different concentrations (0-2000 nm/ml) of LPS and obtained concentrations of TNF-ɑ on each LPS.
The TNF-ɑ concentration increases to about 85pg/ml at 1000 nm/ml of LPS but it drops to 25.73pg/ml at 2000 nm/ml. Anyone knows why TNF-ɑ concentration decrease. I would like to know the reason in detail.
Thank you!!
Hi Daeun, if your experimental conditions for all the LPS groups are identical, it seems your macrophage is exerting an anti-inflammatory response in response to a high LPS dose to counteract the inflammation. Have you checked IL- 10 levels in these groups? Maybe, you will find an answer there. Hope this can help you.
Hi Daeun,
it seems that the high LPS might affect the viability of your cells. In other words it might a supraphysiological dose. It would be better to test different concentrations ~ 1ug per ml.
Good luck
Hi Daeun Roh,
I will highly recommend that you check the viability of the cells as high concentrations of LPS can compromise the viability of the cells; another important question is the time points you used, as the longer you leave the LPS the more cell death will be. On my hands, with human macrophages you should never go higher than 100 ng/ml as this will become physiologically irrelevant. The only rationale for going with higher concentrations is to prove a point.
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