One of the major obstacles for adoptive cell transfer (ACT) of T cells is the loss of effector function and proliferative ability of isolated antigen-specific T cells after prolonged ex vivo expansion. To overcome this issue, induced pluripotent stem cells (iPSCs), which have unlimited proliferation and differentiation potential, can be used to generate a large number of antigen-specific T cells. Here, we describe an efficient differentiation protocol for the generation of cytotoxic CD8+ T cells from human T cell-derived iPSCs (T-iPSCs). The protocol consists of three main steps including differentiation of T-iPSCs toward hematoendothelial progenitors (HEPs), co-culture of HEPs with OP9-DL1 cells, and stimulation of T cell receptor (TCR) signaling to obtain CD8 single-positive (SP) T cells. This culture system is simple and efficient; therefore, will offer a powerful tool for studying T cell development and applications in adoptive immunotherapy.
CITATION STYLE
Suwanpitak, S., Promnakhon, N., Netsrithong, R., & Wattanapanitch, M. (2022). Efficient Generation of iPSC-Derived Hematoendothelial Progenitors and Specification Toward T cell Lineage. In Methods in Molecular Biology (Vol. 2454, pp. 423–442). Humana Press Inc. https://doi.org/10.1007/7651_2021_355
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