An optimized protocol for the retroviral transduction of mouse CD4 T cells

Ekaterina Eremenko; Zoe V Taylor; Bishnu Khand; Shir Zaccai; Angel Porgador; Alon Monsonego

doi:10.1016/j.xpro.2021.100719

An optimized protocol for the retroviral transduction of mouse CD4 T cells

STAR Protoc. 2021 Aug 5;2(3):100719. doi: 10.1016/j.xpro.2021.100719. eCollection 2021 Sep 17.

Authors

Ekaterina Eremenko^{1

2

3

4}, Zoe V Taylor^{1

2

4}, Bishnu Khand^{1

2

4}, Shir Zaccai^{1

2

3

4}, Angel Porgador^{1

2

4}, Alon Monsonego^{1

2

4}

Affiliations

¹ The Shraga Segal Department of Microbiology, Immunology, and Genetics, Faculty of Health Sciences, Ben-Gurion University of the Negev, Beer Sheva 8410501, Israel.
² The Department of Physiology and Cell Biology, Faculty of Health Sciences, Ben-Gurion University of the Negev, Beer Sheva 8410501, Israel.
³ The Department of Life Sciences, Ben-Gurion University of the Negev, Beer Sheva 8410501, Israel.
⁴ The National Institute of Biotechnology in the Negev, Zlotowski Neuroscience Center, and Regenerative Medicine and Stem Cell Research Center, Ben-Gurion University of the Negev, Beer Sheva 8410501, Israel.

Abstract

Transduction of primary T cells has become prominent with the introduction of chimeric antigen receptor T-cell therapy. Although there are many protocols for the transduction of human T cells, it remains a challenge to transduce murine T cells. We present an optimized protocol for the retroviral transduction of murine CD4 T cells, which overcomes major challenges including large-scale production and long-term culturing of transduced cells. The optimized protocol combines high transduction efficiency with a low rate of cell death. For complete details on the use and execution of this protocol, please refer to Eremenko et al., 2019.

Keywords: Cell culture; Cell separation/fractionation; Cell-based Assays; Immunology; Molecular Biology.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Animals
CD4-Positive T-Lymphocytes / immunology
CD4-Positive T-Lymphocytes / metabolism
Flow Cytometry / methods
Genetic Vectors / genetics
Immunotherapy, Adoptive / methods*
Mice
Retroviridae / genetics
Transduction, Genetic / methods*