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

STAR Protoc. 2021 Nov 17;2(4):100956. doi: 10.1016/j.xpro.2021.100956. eCollection 2021 Dec 17.

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).

Keywords: Cancer; Cell Biology; Cell culture; Cell-based Assays; Flow Cytometry/Mass Cytometry; Gene Expression; Immunology; Molecular Bio.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cell Culture Techniques / methods*
  • Cells, Cultured
  • Genetic Vectors / genetics*
  • Humans
  • Immunotherapy, Adoptive
  • Killer Cells, Natural*
  • Lentivirus / genetics*
  • Receptors, Chimeric Antigen / genetics*

Substances

  • Receptors, Chimeric Antigen

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