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

Fingerprint

Cytotoxicity
Natural Killer Cells
Tumors
Assays
Flow cytometry
Color
Cells
Flow Cytometry
Neoplasms
Cell Engineering
U937 Cells
Trypan Blue
Tissue Engineering
Fluorophores
Cell Survival
Tissue engineering
Labeling
fluorexon

Keywords

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

ASJC Scopus subject areas

  • Biotechnology

Cite this

Single-colour flow cytometric assay to determine NK cell-mediated cytotoxicity and viability against non-adherent human tumor cells. / Thakur, Ajit; Zaman, Abeyat; Hummel, Jeff; Jones, Kim; Hortelano, Gonzalo.

In: Biotechnology Letters, Vol. 34, No. 3, 03.2012, p. 447-453.

Research output: Contribution to journalArticle

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