Flow cytometry-based ex vivo murine NK cell cytotoxicity assay

Pamela Wong; Julia A Wagner; Melissa M Berrien-Elliott; Timothy Schappe; Todd A Fehniger

doi:10.1016/j.xpro.2020.100262

Flow cytometry-based ex vivo murine NK cell cytotoxicity assay

STAR Protoc. 2021 Jan 12;2(1):100262. doi: 10.1016/j.xpro.2020.100262. eCollection 2021 Mar 19.

Authors

Pamela Wong¹, Julia A Wagner¹, Melissa M Berrien-Elliott¹, Timothy Schappe¹, Todd A Fehniger¹

Affiliation

¹ Department of Medicine, Division of Oncology, Washington University School of Medicine, St. Louis, MO 63110, USA.

Abstract

Direct killing of diseased cells is a hallmark function of NK cells. This protocol describes a flow-based assay to measure in vivo activated murine NK cells' ability to kill target cells ex vivo. Existing published protocols for assaying ex vivo NK cell killing utilized the radioactive chromium release assay or were designed for human NK cells. This protocol details specifically an ex vivo cytotoxicity assay using primary murine NK cells enriched from splenocytes that were activated in vivo with poly(I:C). For complete details on the use and execution of this protocol, please refer to Wagner et al. (2020).

Keywords: Cell culture; Cell isolation; Cell-based assays; Flow cytometry/mass cytometry; Immunology.

Publication types

Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't

MeSH terms

Animals
Cytotoxicity Tests, Immunologic*
Cytotoxicity, Immunologic*
Flow Cytometry*
Humans
Killer Cells, Natural* / cytology
Killer Cells, Natural* / immunology
Mice
Spleen* / cytology
Spleen* / immunology

Abstract

Publication types

MeSH terms

Grants and funding