IL-6 and TNFa measurement in Caco-2 cells

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Hasan's picture
IL-6 and TNFa measurement in Caco-2 cells

I am trying to investigate the effect of flavonoids on IL-6 and TNF alpha secretion in Caco-2 cells and HRT-18.
should I first stimulate Caco-2 before treatment with flavonoids or without stimulation, and if I should stimulate tumor cell lines, what is the best stimulant is it conA, LPS or IL-1B to release IL-6 and TNF alpha by Caco-2 cells and what is the best concentration for these stimulant and the duration of exposure.
University of jordan.

Kristina Holmberg
Kristina Holmberg's picture
 If your hypothesis is that

 If your hypothesis is that flavonoids will have a therapeutic effect on cytokine relese in cancer cells, then I would suggest that you first stimulate the cells with cytokines etc and then study the effect of the flavonoids.

You need to establish the best condition for each of your cell line and stimulant, both with regard to time and concentration. Once you have done that you pick your optimal concentration and time for your stimulant and then you can do a clasical dose-response expereiment also exploring different exposure times.

Also, to be confident in your results, you should do three independent experiments to establish inter and intra variability.