Kumar Akhilesh, D'Souza Saritha S, Uenishi Gene, Park Mi Ae, Lee Jeong Hee, Slukvin Igor I
Wisconsin National Primate Research Center, University of Wisconsin, Madison, WI 53715, USA.
Department of Cell and Regenerative Biology, School of Medicine and Public Health, University of Wisconsin, Madison, WI 53707, USA.
Bio Protoc. 2020 Jul 5;10(13):e3675. doi: 10.21769/BioProtoc.3675.
Pluripotent stem cells (PSCs) have the potential to provide homogeneous cell populations of T cells that can be grown at a clinical scale and genetically engineered to meet specific clinical needs. OP9-DLL4, a stromal line ectopically expressing the Notch ligand Delta-like 4 (DLL4) is used to support differentiation of PSCs to T-lymphocytes. This article outlines several protocols related to generation of T cells from human and non-human primate (NHP) PSCs, including initial hematopoietic differentiation of PSC on OP9 feeders or defined conditions, followed by coculture of the OP9-DLL4 cells with the PSC-derived hematopoietic progenitors (HPs), leading to efficient differentiation to T lymphocytes. In addition, we describe a protocol for robust T cell generation from hPSCs conditionally expressing ETS1. The presented protocols provide a platform for T cell production for disease modeling and evaluating their use for immunotherapy in large animal models.
多能干细胞（PSC）有潜力提供可在临床规模上培养且经过基因工程改造以满足特定临床需求的T细胞同质细胞群体。OP9-DLL4是一种异位表达Notch配体Delta样4（DLL4）的基质细胞系，用于支持PSC向T淋巴细胞的分化。本文概述了几种从人类和非人类灵长类动物（NHP）PSC生成T细胞的方案，包括在OP9饲养层或特定条件下对PSC进行初始造血分化，随后将OP9-DLL4细胞与PSC衍生的造血祖细胞（HP）共培养，从而高效分化为T淋巴细胞。此外，我们描述了一种从条件性表达ETS1的人PSC中稳健生成T细胞的方案。所提供的方案为疾病建模的T细胞生产以及评估其在大型动物模型中用于免疫治疗提供了一个平台。
