My unstimulated PBMCs (thawed) show a better differentiation and cytokine profile on facs than my E.coli (heat inactivated) stimulated ones (but less than my CD3/28 stimulated positive controls). I have changed media, used different reagents, hoods, autoclaved everything I can think off but not sure why this could be. anyone experienced anything similar or had any ideas for why this might be happening please?
thanks
Hi
There may be contamination in the PBMC culture which you have just thawed. Sometimes contaminants act as stimulants. I have not experienced it myself but I have come across such cases.
I suggest you need to recheck all the vials frozen from the same lot. There is a possibility that the cells may have been contaminated before freezing. This could be one of the possibilities.
Best Wishes.
the only other issue is that I cultured the same cells with inactivated E.coli and I observed less of a response - why would this be if the whole batch was contaminated? should the PBMCs not respond equivocally?
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