Gene expression profile at single cell level of primary regulatory T cells(CD25+CD127- Treg), primary induced regulatory T cells(iTreg), and AMRT treated iPSC-derived CD4+ T cells (iCD4+Treg-like)

We used single cell RNA sequencing (scRNA-seq) to analyze the difference among CD25+CD127-Treg, iTreg, and iCD4+Treg-like. We expanded CD25+CD127-Treg by TCR stimulation in the presense of AS2863619, anti-TNFR2 antibody, and TGFbeta (AMT). We differentiated iTreg from naive CD4+T cells and iCD4+Treg-like from iPSC-derived CD4+T cells in the presence of TCR stimulation, AS2863619, anti-TNFR2 antibody, TGFbeta, and rapamycin (AMRT). CD25+CD127-Treg was isolated from PBMCs by FASC and expanded by TCR stimulation in the presence of AS2863619, anti-TNFR2 antibody, and TGFbeta (AMT) for 14 days. iTreg was differentiated from naïve CD4+ T cells, which were isolated from PBMCs by FACS, in the presence of TCR stimulation, AS2863619, anti-TNFR2 antibody, TGFbeta, and rapamycin (AMRT) for 14 days. iCD4+Treg-like was differentiated from iPSC-derived CD4+T cells in the presence of TCR stimulation, AS2863619, anti-TNFR2 antibody, TGFbeta, and rapamycin (AMRT) for 7 or 14 days.