TY - JOUR
T1 - Genetic Modification of Tumor-Infiltrating Lymphocytes, Peripheral T Cells, and T-Cell Model Cell Lines
AU - Weinstein-Marom, Hadas
AU - Blokon-Kogan, Dayana
AU - Levi-Mann, Maya
AU - Katzman, Chaja
AU - Shalev, Shira
AU - Zaitsev, Masha
AU - Besser, Michal J.
AU - Shapira-Frommer, Ronnie
AU - Gross, Gideon
AU - Itzhaki, Orit
AU - Nissim, Lior
N1 - Publisher Copyright:
© 2024. The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature.
PY - 2024
Y1 - 2024
N2 - 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).
AB - 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).
KW - Adoptive cell therapy
KW - CD8 T cells
KW - Chimeric antigen receptors
KW - Preparation of T cells
KW - Rapid expansion protocol
KW - Retroviral transduction
KW - T cell activation
KW - Tumor-infiltrating lymphocyte
KW - mRNA electroporation
UR - http://www.scopus.com/inward/record.url?scp=85179641182&partnerID=8YFLogxK
U2 - 10.1007/978-1-0716-3593-3_13
DO - 10.1007/978-1-0716-3593-3_13
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C2 - 38070115
AN - SCOPUS:85179641182
SN - 1064-3745
VL - 2748
SP - 167
EP - 186
JO - Methods in Molecular Biology
JF - Methods in Molecular Biology
ER -
