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

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

Research output: Contribution to journal › Article › peer-review

3 Scopus citations

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).

Original language	English
Article number	100262
Journal	STAR Protocols
Volume	2
Issue number	1
DOIs	https://doi.org/10.1016/j.xpro.2020.100262
State	Published - Mar 19 2021

Keywords

Cell culture
Cell isolation
Cell-based assays
Flow cytometry/mass cytometry
Immunology

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10.1016/j.xpro.2020.100262

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title = "Flow cytometry-based ex vivo murine NK cell cytotoxicity assay",

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{\textquoteright} 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",

author = "Pamela Wong and Wagner, {Julia A.} and Berrien-Elliott, {Melissa M.} and Timothy Schappe and Fehniger, {Todd A.}",

note = "Funding Information: This work was supported by grants from the NIH : T32 HL007088 (P.W. and J.A.W.), the Howard Hughes Medical Institute (Medical Fellow Award, J.A.W.), K12 5K12CA167540 (M.M.B.-E.), R01 AI102924 (T.A.F.), R01 CA205239 (T.A.F.), and P50 CA171963 . We acknowledge the use of the Siteman Flow Core for this study. Schema figures were created with Biorender.com . Publisher Copyright: {\textcopyright} 2020",

year = "2021",

month = mar,

day = "19",

doi = "10.1016/j.xpro.2020.100262",

language = "English",

volume = "2",

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AU - Wong, Pamela

AU - Wagner, Julia A.

AU - Berrien-Elliott, Melissa M.

AU - Schappe, Timothy

AU - Fehniger, Todd A.

N1 - Funding Information: This work was supported by grants from the NIH : T32 HL007088 (P.W. and J.A.W.), the Howard Hughes Medical Institute (Medical Fellow Award, J.A.W.), K12 5K12CA167540 (M.M.B.-E.), R01 AI102924 (T.A.F.), R01 CA205239 (T.A.F.), and P50 CA171963 . We acknowledge the use of the Siteman Flow Core for this study. Schema figures were created with Biorender.com . Publisher Copyright: © 2020

PY - 2021/3/19

Y1 - 2021/3/19

N2 - 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).

AB - 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).

KW - Cell culture

KW - Cell isolation

KW - Cell-based assays

KW - Flow cytometry/mass cytometry

KW - Immunology

UR - http://www.scopus.com/inward/record.url?scp=85122830606&partnerID=8YFLogxK

U2 - 10.1016/j.xpro.2020.100262

DO - 10.1016/j.xpro.2020.100262

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AN - SCOPUS:85122830606

SN - 2666-1667

VL - 2

JO - STAR Protocols

JF - STAR Protocols

IS - 1

M1 - 100262

ER -

Flow cytometry-based ex vivo murine NK cell cytotoxicity assay

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Keywords

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