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The development of chimeric antigen receptor (CAR) T cells has been a revolutionary technology for the treatment of relapsed and refractory leukemias and lymphomas. The synthetic CAR molecule redirects T cell function toward tumor surface-expressed antigens through a single-chain variable fragment (scFv) fused to CD3z and intracellular costimulatory domains. Here, we describe a protocol for the generation of CAR T cells using primary mouse T cells and a gammaretroviral vector encoding a CAR transgene. This protocol outlines several transduction and expansion methods based on the use of two transduction enhancers, RetroNectin ® and Vectofusin ® -1, and cell culture systems such as conventional plates or G-Rex ® devices.

Titel:	Expansion and Retroviral Transduction of Primary Murine T Cells for CAR T-Cell Therapy.
Autor/in / Beteiligte Person:	Loos, P ; Short, L ; Savage, G ; Evgin, L
Zeitschrift:	Methods in molecular biology (Clifton, N.J.), Jg. 2748 (2024), S. 41-53
Veröffentlichung:	Totowa, NJ : Humana Press ; <i>Original Publication</i>: Clifton, N.J. : Humana Press,, 2024
Medientyp:	academicJournal
ISSN:	1940-6029 (electronic)
DOI:	10.1007/978-1-0716-3593-3_4
Schlagwort:	Animals Mice Immunotherapy, Adoptive methods Receptors, Antigen, T-Cell genetics Retroviridae genetics T-Lymphocytes Lymphoma
Sonstiges:	Nachgewiesen in: MEDLINE Sprachen: English Publication Type: Journal Article Language: English [Methods Mol Biol] 2024; Vol. 2748, pp. 41-53. MeSH Terms: T-Lymphocytes* ; Lymphoma* ; Animals ; Mice ; Immunotherapy, Adoptive / methods ; Receptors, Antigen, T-Cell / genetics ; Retroviridae / genetics References: Baumann JG, Unutmaz D, Miller MD, Breun SK, Grill SM, Mirro J, Littman DR, Rein A, Kewal Ramani VN (2004) Murine T cells potently restrict human immunodeficiency virus infection. J Virol 78(22):12537–12547. https://doi.org/10.1128/JVI.78.22.12537-12547.2004. (PMID: 10.1128/JVI.78.22.12537-12547.200415507641) ; Tsurutani N, Yasuda J, Yamamoto N, Choi BI, Kadoki M, Iwakura Y (2007) Nuclear import of the preintegration complex is blocked upon infection by human immunodeficiency virus type 1 in mouse cells. J Virol 81(2):677–688. https://doi.org/10.1128/JVI.00870-06. 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(PMID: 10.1016/j.jcyt.2021.11.00435063359) Contributed Indexing: Keywords: Chimeric antigen receptor (CAR); Gamma-retroviral vector; T cell; Transduction; Transfection Substance Nomenclature: 0 (Receptors, Antigen, T-Cell) Entry Date(s): Date Created: 20231209 Date Completed: 20231216 Latest Revision: 20231216 Update Code: 20231218

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Expansion and Retroviral Transduction of Primary Murine T Cells for CAR T-Cell Therapy.