What are the main precautions needed with differentiation of Treg's and TH17 from naive mice T cells?
As both tregs and th17 require TGFb, both may express Foxp3 and Rorgt.
How can I identify them as fully differentiated Tregs or TH17 from mice naive T cells?
Hi,
I think you can use TGF- beta and Il-2 for Treg differentiation, while for Th17 you add Tgf beta plus IL-6, anti IFNg and anti IL-4 if I'm not wrong. At the end of the cell culture you can do a flow cytometry analysis (CD4+CD25+Foxp3+ Treg; CD4+ RORgT expression). You can do some functional assay too, it depends on which is your scientific question.
You can also use CD45RA (at least in humans), to see if the cell has changed the naive phenotype. It is necessary more than CD4 and RORgt to characterizes Th17, you should look for intracellular IL17.
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