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

Ajit Thakur, Abeyat Zaman, Jeff Hummel, Kim Jones, Gonzalo Hortelano

Research output: Contribution to journalArticle

3 Citations (Scopus)

Abstract

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.

Original languageEnglish
Pages (from-to)447-453
Number of pages7
JournalBiotechnology Letters
Volume34
Issue number3
DOIs
Publication statusPublished - Mar 2012
Externally publishedYes

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Keywords

  • Calcein
  • Cell-mediated cytotoxicity
  • Flow cytometry
  • Natural killer cells
  • Viability

ASJC Scopus subject areas

  • Biotechnology

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