I was wondering if culturing mouse BMDM's in 5% FBS vs 10% FBS will have significant differences in cell responses such as gene expression, cytokine production, etc.
Hi,
It is certainly important to control the level of FBS during activation. This is because FBS might contain M-CSF, that inhibits activation, but also LPS-binding proteins, that are required for proper activation (see Dalby, Meth Mol Biol, 1784:111, 2018). We usually differentiate BMDMs in 10% FBS, and perform activation in 2.5% FBS. Note that this might depend on your FBS batch. We have never seen any effect of the FBS concentration change in the absence of activation.
Hope this helps.
Germain
Dear Krishna Chinta, What I remember from experiments with antibody producing cells in vitro is the fact, that not only the amount of FBS was important, but also the serum batch made quite a difference for our results. My advise would be to test a few different batches of FBS as well. Good luck with your work, Willem
FBS concentration can make a large difference in the morphology, physiology, and behavior of your cells, particularly when in monolayer culture. Substrate adherent fibroblasts spread out and their nuclei become flattened, like pancakes. This very much changes their response to ionizing radiations. For historic reasons, monolayer cells were usually grown in 10 (or 15)% FBD, while suspension cultures (as spheroids) were grown in 5% FBS. The lower FBS concentration always produced lower radiosensitivity, irrespective of 2D or 3D culture condition. When medium is isotonically diluted in normal saline, this has the same effect and may explain a significant part (1/2) of why hypoxic starved parts of tumors are more radioresistant.
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