2. Academic Validation
  3. Protocol for the simultaneous activation and lentiviral transduction of primary human T cells with artificial T cell receptors

Protocol for the simultaneous activation and lentiviral transduction of primary human T cells with artificial T cell receptors

  • STAR Protoc. 2025 Mar 21;6(1):103685. doi: 10.1016/j.xpro.2025.103685.
Jordan Yong Ming Tan 1,  Jia Chi Tan 2,  Chuqi Wang 3,  Ling Wu 2,  Nicholas R J Gascoigne 2,  Shruti Bhatt 4
Affiliations

Affiliations

  • 1 Department of Pharmacy & Pharmaceutical Sciences, Faculty of Science, National University of Singapore, 18 Science Drive 2, Singapore 117543, Singapore. Electronic address: jordantanym@u.nus.edu.
  • 2 Department of Microbiology & Immunology, Yong Loo Lin School of Medicine, National University of Singapore, 5 Science Drive 2, Singapore 117545, Singapore; Immunology Translational Research Programme, Yong Loo Lin School of Medicine, National University of Singapore, 5 Science Drive 2, Singapore 117545, Singapore; Cancer Translational Research Programme, Yong Loo Lin School School of Medicine, National University of Singapore, 5 Science Drive 2, Singapore 117545, Singapore.
  • 3 Department of Pharmacy & Pharmaceutical Sciences, Faculty of Science, National University of Singapore, 18 Science Drive 2, Singapore 117543, Singapore.
  • 4 Department of Pharmacy & Pharmaceutical Sciences, Faculty of Science, National University of Singapore, 18 Science Drive 2, Singapore 117543, Singapore. Electronic address: shruti_bhatt@nus.edu.sg.
PMID: 40067824 DOI: 10.1016/j.xpro.2025.103685
Abstract

T cell-based therapies hold immense promise, but their production remains time-consuming and technically complex. Here, we present a protocol that streamlines the activation and lentiviral transduction of primary human T cells with artificial receptors. We describe steps for T Cell Isolation, lentivirus production, and the simultaneous activation/transduction of T cells. By eliminating magnetic T cell activation beads, sequential steps, and lengthy spinoculation, this protocol significantly enhances efficiency, scalability, and accessibility for research and therapeutic applications.

Keywords

Cancer; Cell culture; Gene expression; Immunology; Molecular biology.

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