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
  • 1. Department of Pharmacy & Pharmaceutical Sciences, Faculty of Science, National University of Singapore, 18 Science Drive 2, Singapore 117543, Singapore. Electronic address: [email protected].
  • 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: [email protected].
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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