Abstract
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.
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Acknowledgments
This work was supported by Siriraj Research Fund, Faculty of Medicine Siriraj Hospital, Mahidol University (grant number (IO) R016234002 and R016333015), the Thailand Research Fund (grant no. RSA6280090), the Program Management Unit for Human Resources & Institutional Development, Research and Innovation (grant number B05F630080). NP is supported by the Thailand Research Fund through the Royal Golden Jubilee Ph.D. Program and Faculty of Medicine Siriraj Hospital, Mahidol University (grant no. PHD/0203/2556). RN is supported by the Development and Promotion of Science and Technology Talents Project. MW is supported by Chalermphrakiat Grant, Faculty of Medicine Siriraj Hospital, Mahidol University, Thailand.
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Suwanpitak, S., Promnakhon, N., Netsrithong, R., Wattanapanitch, M. (2021). Efficient Generation of iPSC-Derived Hematoendothelial Progenitors and Specification Toward T cell Lineage. In: Nagy, A., Turksen, K. (eds) Induced Pluripotent Stem (iPS) Cells. Methods in Molecular Biology, vol 2454. Humana, New York, NY. https://doi.org/10.1007/7651_2021_355
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DOI: https://doi.org/10.1007/7651_2021_355
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Publisher Name: Humana, New York, NY
Print ISBN: 978-1-0716-2118-9
Online ISBN: 978-1-0716-2119-6
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