A simple, rapid and sensitive fluorimetric assay for the measurement of cell-mediated cytotoxicity.
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.
Virág
L
L
Department of Pathophysiology, University Medical School of Debrecen, Hungary.
Kerékgyártó
C
C
Fachet
J
J
D000818:Animals; D002455:Cell Division; D003601:Cytotoxicity Tests, Immunologic; D004950:Esterases / Q000378:metabolism; D005260:Female; D005456:Fluorescent Dyes; D006923:Hymecromone / Q000031:analogs & derivatives* / Q000378:metabolism; D066298:In Vitro Techniques; D015979:Killer Cells, Lymphokine-Activated / Q000276:immunology; D051379:Mice; D008806:Mice, Inbred AKR; D008807:Mice, Inbred BALB C; D013050:Spectrometry, Fluorescence
D005456:Fluorescent Dyes; C054268:4-methylumbelliferyl heptanoate; D006923:Hymecromone; D004950:Esterases