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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Bibliographic Details
Main Authors: Ana L. Portillo, Richard Hogg, Ali A. Ashkar
Format: article
Language:EN
Published: Elsevier 2021
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Online Access:https://doaj.org/article/7540abfaed4544eaa9ea2867d0c58adb
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Summary: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).