Literature DB >> 27084117

The modified FACS calcein AM retention assay: A high throughput flow cytometer based method to measure cytotoxicity.

M A Gillissen1, E Yasuda2, G de Jong1, S E Levie2, D Go2, H Spits2, P M van Helden2, M D Hazenberg3.   

Abstract

Current methods to determine cellular cytotoxicity in vitro are hampered by background signals that are caused by auto-fluorescent target and effector cells and by non-specific cell death. We combined and adjusted existing cell viability assays to develop a method that allows for highly reproducible, accurate, single cell analysis by high throughput FACS, in which non-specific cell death is corrected for. In this assay the number of living, calcein AM labeled cells that are green fluorescent are quantified by adding a fixed number of unlabeled calibration beads to the analysis. Using this modified FACS calcein AM retention method, we found EC50 values to be highly reproducible and considerably lower compared to EC50 values obtained by conventional assays, displaying the high sensitivity of this assay.
Copyright © 2016 The Authors. Published by Elsevier B.V. All rights reserved.

Entities:  

Keywords:  ADCC; CDC; Cancer; Cellular cytotoxicity; Monoclonal antibodies

Mesh:

Substances:

Year:  2016        PMID: 27084117     DOI: 10.1016/j.jim.2016.04.002

Source DB:  PubMed          Journal:  J Immunol Methods        ISSN: 0022-1759            Impact factor:   2.303


  9 in total

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Authors:  B Cortjens; E Yasuda; X Yu; K Wagner; Y B Claassen; A Q Bakker; J B M van Woensel; T Beaumont
Journal:  J Virol       Date:  2017-04-28       Impact factor: 5.103

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Journal:  J Immunol       Date:  2018-03-28       Impact factor: 5.422

3.  Patient-derived antibody recognizes a unique CD43 epitope expressed on all AML and has antileukemia activity in mice.

Authors:  Marijn A Gillissen; Greta de Jong; Martijn Kedde; Etsuko Yasuda; Sophie E Levie; Gemma Moiset; Paul J Hensbergen; Arjen Q Bakker; Koen Wagner; Jullien Villaudy; Pauline M van Helden; Hergen Spits; Mette D Hazenberg
Journal:  Blood Adv       Date:  2017-08-18

Review 4.  Comparative analysis of assays to measure CAR T-cell-mediated cytotoxicity.

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Journal:  Nat Protoc       Date:  2021-02-15       Impact factor: 13.491

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Journal:  Organs Chip       Date:  2022-01-26

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Authors:  Luis Romero-Martín; Ferran Tarrés-Freixas; Núria Pedreño-López; Maria L Rodríguez de la Concepción; Francesc Cunyat; Dennis Hartigan-O'Connor; Jorge Carrillo; Beatriz Mothe; Julià Blanco; Marta Ruiz-Riol; Christian Brander; Alex Olvera
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  9 in total

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