Protocol for in vitro isolation, induction, expansion, and determination of human natural regulatory T cells and induced regulatory T cells

Regulatory T cells (Tregs) can inhibit the occurrence of autoimmune diseases and increase the activation threshold of the immune response. Here, we present schemes for the isolation and culture of natural human regulatory T cells (nTregs) and in vitro-induced Tregs (iTregs). Appropriate concentrations of TGF-β, IL-2, retinoic acid (atRA), and rapamycin were used to promote proliferation to meet sample needs in basic research, especially in technologies such as sequencing. For complete details on the use and execution of this protocol, please refer to Lu et al. (2014a) and Gu et al. (2022).