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Background  T cell development occurs within the highly specialized thymus. Cytotoxic CD8 T cells are critical in adaptive immunity by targeting virally infected or tumor cells. In this study, we addressed whether functional CD8 T cells can be generated fully  in vitro  using human umbilical cord blood (UCB) hematopoietic stem cells (HSCs) in coculture with OP9-DL1 cells.    Results  HSC/OP9-DL1 cocultures supported the differentiation of CD8 T cells, which were TCR/CD3 hi  CD27 hi  CD1a neg  and thus phenotypically resembled mature functional CD8 single positive thymocytes. These  in vitro -generated T cells also appeared to be conventional CD8 cells, as they expressed high levels of  Eomes  and low levels of  Plzf , albeit not identical to  ex vivo  UCB CD8 T cells. Consistent with the phenotypic and molecular characterization, upon TCR-stimulation,  in vitro -generated CD8 T cells proliferated, expressed activation markers (MHC-II, CD25, CD38), secreted IFN-γ and expressed Granzyme B, a cytotoxic T-cell effector molecule.    Conclusion  Taken together, the ability to direct human hematopoietic stem cell or T-progenitor cells towards a mature functional phenotype raises the possibility of establishing cell-based treatments for T-immunodeficiencies by rapidly restoring CD8 effector function, thereby mitigating the risks associated with opportunistic infections.

Human CD8 T cells generated in vitro from hematopoietic stem cells are functionally mature