Suwanpitak S.Promnakhon N.Netsrithong R.Wattanapanitch M.Mahidol University2023-06-182023-06-182022-01-01Methods in Molecular Biology Vol.2454 (2022) , 423-44210643745https://repository.li.mahidol.ac.th/handle/123456789/83901One 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.Biochemistry, Genetics and Molecular BiologyBook ChapterSCOPUS10.1007/7651_2021_3552-s2.0-851324557521940602933755900