Siriraj Center for Regenerative Medicine, Research Department, Faculty of Medicine, Siriraj Hospital, Mahidol University, Bangkok, Thailand.
Department of Immunology, Faculty of Medicine, Siriraj Hospital, Mahidol University, Bangkok, Thailand.
Methods Mol Biol. 2022;2454:423-442. doi: 10.1007/7651_2021_355.
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.
过继细胞转移（ACT）的主要障碍之一是分离的抗原特异性 T 细胞在体外长期扩增后效应功能和增殖能力的丧失。为了克服这个问题，可以使用诱导多能干细胞（iPSCs），其具有无限增殖和分化潜能，从而产生大量的抗原特异性 T 细胞。在这里，我们描述了一种从人 T 细胞来源的诱导多能干细胞（T-iPSCs）产生细胞毒性 CD8 T 细胞的有效分化方案。该方案包括三个主要步骤，包括 T-iPSCs 向造血内皮祖细胞（HEPs）的分化、HEPs 与 OP9-DL1 细胞的共培养以及 T 细胞受体（TCR）信号的刺激以获得 CD8 单阳性（SP）T 细胞。该培养系统简单高效；因此，将为研究 T 细胞发育和过继免疫治疗中的应用提供有力工具。
