I started to work with C8-D1A cells, mouse astrocyte cell line.
We are trying to induce inflammation in cells using LPS, but treatment with LPS does not seem to induce inflammation.
I also purchased new LPS and tried various adjustments to the LPS concentration(~2uM/ml) and treatment time(~48h), but there was no significant difference.
Is there a way to tell if this is due to a high level of basal inflammation in the cells?
Or people who have had similar experiences ask for advice
Hi, can you provide the amount of LPS you tesed in EU/ml or µg/ml? And furhtermore, what is your read-out for the inflammatory phenotype?
Hi
I tested LPS concentrations from 100ng/ml to 2ug/ml.
And I checked the mRNA expression of inflammatory cytokines, including TNF-a, IL-1B, and IL-6.
You should try to measure ROS in your cells using some assay kit like DCFDA. You should obtain a big ROS increase compared to non-treated sample and therefore prove that LPS treatment works and induce inflammation
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