Our lab has been working with fresh PBMC for decades. We separate them from buffy coats and seed them, incunate them overnight, then we treat them with inducers (SEB, LPS and others) and measure inflammatory cytokine output.
Lately I've noticed that our negative control which receives no treatment whatsoever also secretes cytokines (we've seen this with IL6 and IL8), especially in the first hour after treatment (we measure 0h, 1h, 2h and a few more time points). Sometimes it levels off after the first hour, but sometimes the cytokine levels keep climbing just like the PBMC that did receive treatment with an inducer.
What could be the cause of this? And more importantly, how do I avoid it?
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