Hi all,
I am curious that why in some methods, people add IL1b and IL23 on top of the TGFB and IL6 for Th17 cells differentiation in vitro?
Also, the percentage of Th17 after in vitro differentiation is only around 30 percent. Why is it not 100%? What cell types are the other 70% cells? What's the principle behind this phenomenon?
Any suggestion or reference will be appreciated,
Thanks
I think the answer you are looking for is included in this article below
Article Insights into the mechanisms of Th17 differentiation and the...
I think (according to several papers I've read) that 100% is utopistic. 30% is quite good but maybe you could improve it with addition of an anti-IFN-g antibody to block unwanted TH1 differentiation which would impair Th17-skewing (also anti-IL-4 could be added).
IL-23 is not absolutely required for differentiation but rather stabilizes the phenotype and can increase the yield.
For IL-1b I'm not sure (we don't use it for Th17 differentiation), but I think to recall that IL-1b is way more important for human Th17.
Also, if you want to further increase Th17 yield (although a bit artificially), you could increase the amount of NaCl in your medium since it is known that higher amount of NaCl fosters Th17 differentiation.
Hope this helps.
Hi Xingchen,
You can also add anti-IL-2 to you culture for closer to 70% IL-17A+ cells. But it is likely the a-IL-2 is having more of a negative impact on the IL-17A (-) CD4 cells, which enables the higher frequency.
I only can get 5% of il17 A + th17 T cells after differentaition from naive cd4 + T cells , could you show me your protocol , I have been trying this for half a year
Xingchen Dong
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