Can I use multiple donors for T cell assays? Will mixing cause unwanted stimulation?Peripheral blood mononuclear cells will be used to track proliferation via CFSE in vitro. Stimulation via PMA to investigate mechanisms of immune dampening when co-cultured with MSC's.
Everytime I did human T cell assays I used each donor as a individual sample (triplicates from each donor) and ideally the more donors the better. Specially since humans are not isogenic and cells from different donors will respond differently. If you mix cells you will have an average from all donors, in which some respond better than others. In this case you will lose data and probably will be asked by the reviewer when you try to publish your article.
Even with identical MHC haplotype, differences in minor antigens will cause cross stimulation. identical haplotypes are present only in identical twins.
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