Production of human CAR-NK cells with lentiviral vectors and functional assessment in vitro

Summary: Although natural killer (NK) cells have become a promising immune effector cell for chimeric antigen receptor (CAR)-based therapy, generating human CAR-NK cells with high transgene efficiency has been challenging. In this protocol, we describe how to generate CAR-NK cells with transduction...

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Autores principales:	Ana L. Portillo, Richard Hogg, Ali A. Ashkar
Formato:	article
Lenguaje:	EN
Publicado:	Elsevier 2021
Materias:	Cancer Cell Biology Cell culture Cell-based Assays Flow Cytometry/Mass Cytometry Gene Expression Science (General) Q1-390
Acceso en línea:	https://doaj.org/article/7540abfaed4544eaa9ea2867d0c58adb
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spelling	oai:doaj.org-article:7540abfaed4544eaa9ea2867d0c58adb2021-11-18T04:51:41ZProduction of human CAR-NK cells with lentiviral vectors and functional assessment in vitro2666-166710.1016/j.xpro.2021.100956https://doaj.org/article/7540abfaed4544eaa9ea2867d0c58adb2021-12-01T00:00:00Zhttp://www.sciencedirect.com/science/article/pii/S2666166721006626https://doaj.org/toc/2666-1667Summary: Although natural killer (NK) cells have become a promising immune effector cell for chimeric antigen receptor (CAR)-based therapy, generating human CAR-NK cells with high transgene efficiency has been challenging. In this protocol, we describe how to generate CAR-NK cells with transduction efficiencies >15% from healthy donor ex vivo expanded NK cells using third generation lentiviral vectors (LVs). We also show how to assess CAR-NK cell anti-tumor function in vitro using a flow cytometry-based killing assay.For complete details on the use and execution of this protocol, please refer to Portillo et al. (2021).Ana L. PortilloRichard HoggAli A. AshkarElsevierarticleCancerCell BiologyCell cultureCell-based AssaysFlow Cytometry/Mass CytometryGene ExpressionScience (General)Q1-390ENSTAR Protocols, Vol 2, Iss 4, Pp 100956- (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Cancer Cell Biology Cell culture Cell-based Assays Flow Cytometry/Mass Cytometry Gene Expression Science (General) Q1-390
spellingShingle	Cancer Cell Biology Cell culture Cell-based Assays Flow Cytometry/Mass Cytometry Gene Expression Science (General) Q1-390 Ana L. Portillo Richard Hogg Ali A. Ashkar Production of human CAR-NK cells with lentiviral vectors and functional assessment in vitro
description	Summary: Although natural killer (NK) cells have become a promising immune effector cell for chimeric antigen receptor (CAR)-based therapy, generating human CAR-NK cells with high transgene efficiency has been challenging. In this protocol, we describe how to generate CAR-NK cells with transduction efficiencies >15% from healthy donor ex vivo expanded NK cells using third generation lentiviral vectors (LVs). We also show how to assess CAR-NK cell anti-tumor function in vitro using a flow cytometry-based killing assay.For complete details on the use and execution of this protocol, please refer to Portillo et al. (2021).
format	article
author	Ana L. Portillo Richard Hogg Ali A. Ashkar
author_facet	Ana L. Portillo Richard Hogg Ali A. Ashkar
author_sort	Ana L. Portillo
title	Production of human CAR-NK cells with lentiviral vectors and functional assessment in vitro
title_short	Production of human CAR-NK cells with lentiviral vectors and functional assessment in vitro
title_full	Production of human CAR-NK cells with lentiviral vectors and functional assessment in vitro
title_fullStr	Production of human CAR-NK cells with lentiviral vectors and functional assessment in vitro
title_full_unstemmed	Production of human CAR-NK cells with lentiviral vectors and functional assessment in vitro
title_sort	production of human car-nk cells with lentiviral vectors and functional assessment in vitro
publisher	Elsevier
publishDate	2021
url	https://doaj.org/article/7540abfaed4544eaa9ea2867d0c58adb
work_keys_str_mv	AT analportillo productionofhumancarnkcellswithlentiviralvectorsandfunctionalassessmentinvitro AT richardhogg productionofhumancarnkcellswithlentiviralvectorsandfunctionalassessmentinvitro AT aliaashkar productionofhumancarnkcellswithlentiviralvectorsandfunctionalassessmentinvitro
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Production of human CAR-NK cells with lentiviral vectors and functional assessment in vitro

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