A Serum- and Feeder-Free System to Generate CD4 and Regulatory T Cells from Human iPSCs

While feeder-free generation of clinical grade iPSC-derived CD8 T cells has been achieved, differentiation of iPSC-derived CD4sp and regulatory T cells requires mouse stromal cells in an artificial thymic organoid. We developed a serum- and feeder-free differentiation process suitable for large-scale production and used scRNAseq to study the cell identity of the iPSC-derived cells and compared their transcriptional profile to primary human cells. RNA from human iPSC-Tregs, iPSC-CD4sp, primary Tregs, induced Tregs and primary Tconv was extracted following differentiation (iPSC-Tregs and induced Tregs) or isolation and culture (primary Tregs and Tconv). Human primary cells were isolated from leukapheresis.