I am establishing a mo-DC protocol, and there is too many option in regards of maturation. I would like some help deciding. Since later in downstream applications I would be combining IFNg, I would like to include this cytokine in the protocol. However, I've seen that in order to truly create maDC i should activate the other canonical pathways. That is why I though of using TNFa and LPS. Before jumping and trying it out, I was wondering if anyone has experience with this type of cocktail or IFNg+TNFa.
Thanks
Hi Ramon,
there a quite a few maturation cocktails already established. I have tried out four of five of them.
This paper describes three cocktails and also includes IFNgamma: https://www.ncbi.nlm.nih.gov/pubmed/19342965
In my last project in ended up using a combination of PGE2, IFNgamma, TNF and IL-1beta. I also know quite a few labs use only Poly I:C or only LPS (I had good results with both) - but I guess that also depends on your personal preference and what you would like to do with your mDCs.
Hope that helps you.
Cheers, Lisa
Thank you Lisa and Cemsel, it seems like the INFg +LPS is a safer and more studied option. I just cannot find anything that only uses IFNg + TNFa, I though that combining these two molecules will recapitulate some of the intracellular+extracellular insult. Thanks
Hi
Try LPS + IFNg + Poly I:C for max TLR stimulation. I would not try TNFa if you want a good TH1 output from your DC. TNFa used to be used along with IL-6 and PGE2, but now you don't see them used as they can create a DC2 profile to Tregs.
Bruce
Bruce! Thanks for such informative response. As I was waiting for reagents I tried the INFg and TNFa and it does indeed have something like a mature tolDC or DC2 profile. I would never though that TNFa was possible of doing that. Thank you
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