Greeting everyone,
Recently, our lab want to create an in vitro inflammatory cell model by recombinant TNF-a on either 3T3-L1 or HL-1 (cardiomyocytes). I read several articles and many methods showed they used serum-free medium when they added rTNF-a to the cell. However, I can not find any evidences that show FBS would influence the function of TNF-a in the cell. I think cells will under a starvation condition further trigger more stress signals and influence the result. Does anyone knows why those protocols remove FBS when they add TNF-a into the cell?
Thank you for your patience and response.
Best regards,
Hello Bo-Yao Wen
FBS-supplemented medium carries a certain amount of serum components which could potentially trigger unwanted stimulation that could influence your end results. I agree with you that cells under starvation could induce unique stress signaling pathways that could have an effect on the results.
So, I would suggest, you grow cells in serum-containing medium until they reach the required confluency. Then change the medium to serum-free medium, keep the cells in serum free-medium for a few hours, and then add rTNF-a to the cells.
If you feel that serum starvation could induce various responses that can interfere with experimental results, as serum starvation has been shown to cause cells to undergo apoptosis and autophagy, you may perform the treatment with medium containing low serum, say 0.5 -1%.
Best.
Thank you Malcolm Nobre .
That is what I assume FBS contains various ingredients, including Interleukins. But I am not sure whether those would affect the outcome significantly. (even more than starvation?)
Your suggestion is great. I will try to test the cells with a low serum to see how it goes.
Thank you again for your kind answer.
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