Hey Everyone,
I'm trying to plan out a DC T-cell co culture and I'm not sure if i'll need to add IL-2 to my culture to support my T cells during my incubation (4-5 days for proliferation assay). If my DCs have been previously conditioned for 24 hours by intestinal epithelial cell media (that has been previously stimulated with Flagellin=TLR5 agonist), can I rely on the DCs sending the necessary signals to support T cell growth and proliferation? Im certain CD80/86 will be upregulated-- I'm just worried without the addition of IL-2, my T cells will not be happy.
Any thoughts?
One note: My Flagellin has been removed from the media, so no need to worry about residual flagellin interacting with the DCs and T cells.
William D Rees why are you avoiding IL-2 in your co-culture? Is there a specific reason? in general it is recommended to use IL-2 to keep the T cells happy (~ 50U/ml).
Thanks for the response. I haven’t thought of a reason to not add it. I didnt know if it was required and if I dont have to add IL-2, I would rather not.
You don"t need to add IL-2 if you keep proportion of 5-10 T cells to 1 DC.
Ozkan Gelincik, maybe you can help me: Im doing coculture of mo-DC and TCD4 cells (1:4), and I'm questioning if the mature mo-DC (LPS maturated) are capable to induce Tcell differentiation. If I use the IL-2 in the protocol, this are going to be an interference on my results, isn't? I'm going to quantify the IL-2 intracelular concentration by FACS.
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