A simple, rapid and sensitive fluorimetric assay for the measurement of cell-mediated cytotoxicity

László Virág, Csilla Kerékgyártó, József Fachet

Research output: Contribution to journalArticle

28 Citations (Scopus)

Abstract

A fluorimetric method using 4-methylumbelliferyl heptanoate (MUH) has been developed for detecting cell-mediated cytotoxicity and cell proliferation. The assay is based on the hydrolysis of the fluorochrome (MUH) by intracellular esterases of viable cells resulting in the production of highly fluorescent 4-methylumbelliferone that can be measured in a microplate fluorimeter. Because of a similarity to the principle of the widely used colorimetric MTT assay, a comparison was made between the two assays when measuring cell proliferation and LAK cell cytotoxicity to different target cell types. The results have shown that the MUH assay represents a method for evaluating both cell-mediated cytotoxicity and cell proliferation which is completely comparable to the MTT method. The rapidity of the new cytotoxicity assay, 5 h in contrast to 9 h for the MTT assay, its applicability to both adherently and nonadherently growing target cells and its high accuracy due to the avoidance of centrifugation steps make this method a serious contender for replacing conventional radioactive techniques.

Original languageEnglish
Pages (from-to)199-208
Number of pages10
JournalJournal of Immunological Methods
Volume185
Issue number2
DOIs
Publication statusPublished - 1995

Keywords

  • 4-Methylumbelliferyl heptanoate
  • Cell-mediated cytotoxicity
  • Fluorimetric assay
  • Lymphokine-activated killer cell
  • Tumor cell line

ASJC Scopus subject areas

  • Immunology and Allergy
  • Immunology

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