Single-colour flow cytometric assay to determine NK cell-mediated cytotoxicity and viability against non-adherent human tumor cells

被引：0
作者：
Ajit Thakur
Abeyat Zaman
Jeff Hummel
Kim Jones
Gonzalo Hortelano
机构：
[1] McMaster University,School of Biomedical Engineering
[2] McMaster University,Bachelor of Health Sciences Program
[3] McMaster University,Centre for Gene Therapeutics
[4] McMaster University,Department of Chemical Engineering
[5] McMaster University,Department of Pathology & Molecular Medicine
来源：
Biotechnology Letters | 2012年 / 34卷
关键词：
Calcein; Cell-mediated cytotoxicity; Flow cytometry; Natural killer cells; Viability;
D O I：
暂无
中图分类号：
学科分类号：
摘要：
A flow cytometry-based cytotoxicity (FCC) assay was developed using a single fluorophore, calcein-acetoxymethyl diacetylester (calcein-AM), to measure NK cell-mediated cytotoxicity. Non-adherent human K562 and U937 target cells were individually labelled with calcein-AM and co-incubated with effector NK cells to measure calcein loss, and therefore calculate target cell cytotoxicity. This FCC assay also provided a measure of sample viability. Notably, cell viability measured by traditional calcein/7-amino-actinomycin D (7-AAD) double labelling and Trypan Blue methods were comparable to the viability calculated using calcein-loss FCC. This FCC assay may also be used with various effector and target cell types and as a multi-parameter tool to measure viability and immunophenotype cells for tissue engineering purposes.
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页码：447 / 453
页数：6
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