I am using the RAW 264.7 cell line to do a drug release study. They are activated with 10 ug/mL LPS. When changing the medium after two or three days, do I need to add LPS to the new medium as well? I plan to do the release study over seven days.
So, the macrophages should still be activated for several days once the medium with LPS is removed, correct?
You can also parallelly measure macrophages activation markers (phenotypic or cytokines), which will confirm you that they are activated throughout your experimental period (e.g. 7 days). Alternatively, one can rule out the effect of LPS addition during medium changing in two or three days interval. For instance, one experiment with only first LPS stimulation of the macrophages and then replacing medium with non-LPS containing medium. The second experiment would be replacing the medium with LPS-containing medium (with constant concentration) and finally you will see the dynamic of your analyte (interest) in these two experimental design.
Good luck!
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