Using HUVEC cells: My media has low FBS (2%). I wanted to look at cytokine production after LPS stimulation without removing FBS from the media (because it's possible that it's removal may stress the cells also). I have read that some cytokines are affected in other cells lines if FBS is present during LPS incubation vs. if it is absent. Has anyone had any issues with leave the FBS in while incubating with LPS as far as cytokine levels?
Dear Holly,
Whether or not FBS supplementation will affect response to LPS stimulation will most likely depend on the type of FBS you're using. One type of FBS may inhibit LPS effects while another one may boost it. Even if you use exactly the same FBS reference from the same company, you may have substantial differences from one batch of FBS to another.
This well-known reproducibility problem with FBS has recently been highlighted in a Nature comment.
Overall, the (only) question you can ask is whether the FBS you are using right-now has an impact on your LPS stimulation. It is also quite common to test a dozen of different FBS batches for a given assay, and to subsequently order a large amount of the FBS that works best to avoid facing differences afterwards that would be due to different FBS batches.
Hope this helps.
https://www.nature.com/nature/journal/v537/n7620/full/537433a.html
If you are worried about viability, you can also use 0.02% BSA supplemented medium. This helps maintain an osmotic balance in the absence of serum.
Dear Holly,
TLR signaling involves production of reactive oxygen species. FCS contains glutathion and cystein. Thus, FCS reduces oxidative signaling resulting in altered gene (cytokine) expression. Therefore, I agree with Julien. You have to test various batches of FCS to find the optimal batch for your cell culture. However, some people dialyse the FCS to dilute cysteine and gluthation out.
Best,
Karsten
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