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MTT colorimetric assay for testing macrophage cytotoxic activity in vitro
M Ferrari1, M C Fornasiero, A M Isetta
1Immunology Line, Farmitalia Carlo Erba R&D-Erbamont Group, Nerviano, Italy.
Journal of Immunological Methods
|August 7, 1990
Summary
The MTT assay quantifies macrophage cytotoxicity by measuring formazan production, offering a sensitive, reproducible, and rapid alternative to radioisotopic methods for evaluating cell survival.
Area of Science:
- Immunology
- Cell Biology
Background:
- The MTT assay measures cell survival and proliferation.
- It has been adapted to quantify macrophage-mediated cytotoxicity.
Purpose of the Study:
- To evaluate the MTT assay's effectiveness in quantifying macrophage cytotoxicity.
- To compare the MTT assay with radioisotopic methods like 3H-TdR uptake and release assays.
Main Methods:
- Used peritoneal macrophages as effectors and TU5/L929 cell lines as targets in a 48-hour cytotoxicity test.
- Quantified MTT reduction to formazan spectrophotometrically at 570 nm.
- Compared MTT assay results with 3H-TdR uptake and release assays.
Main Results:
- Demonstrated a linear relationship between formazan generated and viable target cells.
- MVE2-activated macrophages showed dose-dependent inhibition of target cell survival.
- MTT assay showed comparable or superior sensitivity to radioisotopic methods.
Conclusions:
- The MTT assay is a sensitive, reproducible, and rapid alternative to radioisotopic methods for quantifying macrophage cytotoxicity.
- It eliminates the need for radioactive compounds and is easy to perform.
- The method is suitable for actively growing in vitro targets.