Retroviral transduction of murine primary T lymphocytes. Academic Article uri icon

Overview

abstract

  • In comparison to human T cells, efficient retroviral gene transfer and subsequent expansion of murine primary T cells is more difficult to achieve. Herein, we describe an optimized gene transfer protocol utilizing an ecotropic viral vector to transduce primary murine T cells activated with magnetic beads coated with agonistic anti-CD3 and CD28 antibodies. Activated T cells are subsequently centrifuged (spinoculated) on RetroNectin-coated tissue culture plates in the context of retroviral supernatant. Variables found to be critical to high gene transfer and subsequent efficient T cell expansion included CD3/CD28 magnetic bead to cell ratio, time from T cell activation to initial spinoculation, frequency of T cell spin-oculation, interleukin-2 concentration in the medium, and the initial purity of the T cell preparation.

publication date

  • January 1, 2009

Research

keywords

  • Retroviridae
  • T-Lymphocytes
  • Transduction, Genetic

Identity

PubMed Central ID

  • PMC5003426

Scopus Document Identifier

  • 60749131428

Digital Object Identifier (DOI)

  • 10.1007/978-1-59745-409-4_7

PubMed ID

  • 19110621

Additional Document Info

volume

  • 506