Genetic Modification of Tumor-Infiltrating Lymphocytes, Peripheral T Cells, and T-Cell Model Cell Lines

Hadas Weinstein-Marom, Dayana Blokon-Kogan, Maya Levi-Mann, Chaja Katzman, Shira Shalev, Masha Zaitsev, Michal J. Besser, Ronnie Shapira-Frommer, Gideon Gross, Orit Itzhaki, Lior Nissim

Research output: Contribution to journalArticlepeer-review

Abstract

Genetic modification of tumor-infiltrating lymphocytes (TILs) or circulating T cells has become an important avenue in cancer therapy. Here we describe a comprehensive method for establishing and expanding TIL cultures and genetically modifying them with a gene of interest (GOI) via retroviral transduction or mRNA transfection. The method includes all the important steps starting with TIL extraction from tumors through to the maintenance of the genetically modified TILs. The protocol includes instructions for retroviral transduction and mRNA transfection of circulating T cells or T-cell lines. The GOIs most commonly introduced into the target cells are chimeric antigen receptors (CARs); genetic adjuvants, such as membrane-bound interleukins; and antitumor T-cell receptors (TCRs).

Original languageAmerican English
Pages (from-to)167-186
Number of pages20
JournalMethods in Molecular Biology
Volume2748
DOIs
StatePublished - 2024

Bibliographical note

Publisher Copyright:
© 2024. The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature.

Keywords

  • Adoptive cell therapy
  • CD8 T cells
  • Chimeric antigen receptors
  • Preparation of T cells
  • Rapid expansion protocol
  • Retroviral transduction
  • T cell activation
  • Tumor-infiltrating lymphocyte
  • mRNA electroporation

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