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

ABSTRACT: 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).

SUBMITTER: Portillo AL

PROVIDER: S-EPMC8605094 | biostudies-literature | 2021 Dec

REPOSITORIES: biostudies-literature

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

Portillo Ana L AL Hogg Richard R Ashkar Ali A AA

STAR protocols 20211117 4

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 cy ...[more]

PMID: 34825217

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