Organoid-Induced Differentiation of Conventional T Cells from Human Pluripotent Stem Cells

Summary The ability to generate T cells from pluripotent stem cells (PSCs) has the potential to transform autologous T cell immunotherapy by facilitating universal, off-the-shelf cell products. However, differentiation of human PSCs into mature, conventional T cells has been challenging with existing methods. We report that a continuous 3D organoid system induced an orderly sequence of commitment and differentiation from PSC-derived embryonic mesoderm through hematopoietic specification and efficient terminal differentiation to naive CD3 + CD8αβ + and CD3 + CD4 + conventional T cells with a diverse T cell receptor (TCR) repertoire. Introduction of an MHC class I-restricted TCR in PSCs produced naive, antigen-specific CD8αβ + T cells that lacked endogenous TCR expression and showed anti-tumor efficacy in vitro and in vivo . Functional assays and RNA sequencing aligned PSC-derived T cells with primary naive CD8 + T cells. The PSC-artificial thymic organoid (ATO) system presented here is an efficient platform for generating functional, mature T cells from human PSCs.