Generation of T cells from Human and Nonhuman Primate Pluripotent Stem Cells

Akhilesh Kumar; Saritha S D'Souza; Gene Uenishi; Mi Ae Park; Jeong Hee Lee; Igor I Slukvin

doi:10.21769/BioProtoc.3675

Generation of T cells from Human and Nonhuman Primate Pluripotent Stem Cells

Bio Protoc. 2020 Jul 5;10(13):e3675. doi: 10.21769/BioProtoc.3675.

Authors

Akhilesh Kumar¹, Saritha S D'Souza¹, Gene Uenishi¹, Mi Ae Park¹, Jeong Hee Lee¹, Igor I Slukvin^{1

2

3}

Affiliations

¹ Wisconsin National Primate Research Center, University of Wisconsin, Madison, WI 53715, USA.
² Department of Cell and Regenerative Biology, School of Medicine and Public Health, University of Wisconsin, Madison, WI 53707, USA.
³ Department of Pathology and Laboratory Medicine, University of Wisconsin, Madison, WI 53792, USA.

Abstract

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.

Keywords: Hematopoietic differentiation; Hematopoietic progenitor; Hemogenic endothelium; Human embryonic stem cells; Human pluripotent stem cells; Non-human primate pluripotent stem cells; T cells.

Abstract

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