Work

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

Year: 2025

Type: article

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. • Steps for transfecting HEK293T cells with DNA plasmids to produce lentiviral particles • Instructions for the activation and lentiviral transduction of primary human T cells • Guidance on the culture and expansion of transduced T cells Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics. 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.

Source: STAR Protocols

DOI: 10.1016/j.xpro.2025.103685