I usually perform short-term treatments (6h) and so I use serum free medium, but in a new protocol I have to treat for 5 days Caco-2 cells grown on Transwell membranes. In this case should I use medium with 10% FBS or reduced percentage of FBS instead of serum free? Cells are treated with TNF, LPS and IL-1b, other secreted cytokines are measured in the medium by ELISA assay. Can the FBS in the medium interfere with the stimuli or with the subsequent ELISA assay?
Thanks for your answers.
I regularly use LPS treatments in serum enriched media and it works fine (no problems--- though the treatment is just 24h). I cannot speak for the other assays you plan to run. Best of luck!
Elliot
Dear Researcher,
FBS is needs to grow the cells. This is growth factor. As your reaction is for 5 days. It is practically impossible to grow cells without FBS. So you can perform ELISA of only 10% FBS containing Medium simultaneously along with your test sample and during the calculation subtract these value from test sample.
Than you very much! Since my protocol involves medium changing every day, I'll try serum deprivation only on the last one, just to avoid possible interference with downstream assays.
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