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. Epub 2025 Mar 10.

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.

MeSH terms

  • Genetic Vectors / genetics
  • HEK293 Cells
  • Humans
  • Lentivirus* / genetics
  • Lymphocyte Activation
  • Receptors, Antigen, T-Cell* / genetics
  • Receptors, Antigen, T-Cell* / metabolism
  • Receptors, Artificial* / genetics
  • Receptors, Artificial* / metabolism
  • T-Lymphocytes* / cytology
  • T-Lymphocytes* / immunology
  • T-Lymphocytes* / metabolism
  • Transduction, Genetic* / methods

Substances

  • Receptors, Antigen, T-Cell
  • Receptors, Artificial