Currently, T cell-based immunotherapies generally rely on autologous T cells, limiting their use 4, 5. T cells are an important cellular component of the immune system that can be harnessed for both regenerative and immunotherapeutic approaches 1, 2, 3. DL4-μbeads provide a simple and robust platform to both study human T cell development and facilitate the development of engineered T cell therapies from renewable sources. Furthermore, the adoptive transfer of CD34 +CD7 + proT cells into immunodeficient mice demonstrates efficient thymic engraftment and functional maturation of peripheral T cells. Single-cell RNA sequencing of human PSC-derived proT cells reveals a transcriptional profile similar to the earliest thymocytes found in the embryonic and fetal thymus. DL4-μbeads, along with lymphopoietic cytokines, induce an ordered sequence of differentiation from CD34 + cells to CD34 +CD7 +CD5 + proT cells to CD3 +αβ T cells. Here, we report a stromal cell-free, microbead-based approach that supports the efficient in vitro development of both human progenitor T (proT) cells and T-lineage cells from CD34 +cells sourced from cord blood, GCSF-mobilized peripheral blood, and pluripotent stem cells (PSCs). An unmet challenge in the field is the development of a clinically relevant system that is readily scalable to generate large numbers of T-lineage cells from hematopoietic stem/progenitor cells (HSPCs). T cells are pivotal effectors of the immune system and can be harnessed as therapeutics for regenerative medicine and cancer immunotherapy.
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