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Semiautomated colorimetric assay for in vitro screening of anticancer compounds
1Medical Products Department, E.I. Du Pont de Nemours and Company, Glenolden, PA 19036.
Abstract:
An in vitro tetrazolium dye (MTT) reduction technique was modified and evaluated for use in the large-scale screening of anticancer compounds by examining the activity of ten clinically used drugs against 16 different human and murine cell populations. Cell populations included colon and mammary adenocarcinomas, melanomas, leukemias, and freshly isolated normal cells. Cell lines were grown in microtiter plates for 18-20 hours prior to a 72-hour continuous exposure to the drugs. Cultures were initiated at cell densities which maximized both the difference in dye reduction and the number of cell doublings between the beginning and end of the drug exposure period. Drug potency, expressed as the 50% inhibitory concentration (IC50), was comparable whether the effect on cell doublings or dye reduction was determined. There was good agreement between this method and the more labor-intensive, conventional method of counting trypan blue dye-excluding cells in a hemacytometer. Implemented as a large-scale, high-capacity system, our adaptation of the MTT technique is a rapid, sensitive, reproducible first-line screening device for detecting anticancer compounds with cytostatic or cytocidal activity.
Insights
A modified tetrazolium dye (MTT) assay offers a rapid, sensitive method for screening anticancer compounds. This in vitro technique effectively identifies cytostatic or cytocidal drug activity across diverse cell populations.
Area of Science:
- Pharmacology and Toxicology
- Cell Biology and Cancer Research
Background:
- Accurate and efficient screening of potential anticancer compounds is crucial for drug discovery.
- Traditional cell-based assays for drug screening can be labor-intensive and time-consuming.
Purpose of the Study:
- To modify and evaluate the tetrazolium dye (MTT) reduction assay for large-scale anticancer compound screening.
- To assess the assay's sensitivity, reproducibility, and comparability to conventional methods.
Main Methods:
- Modified MTT assay applied to 16 human and murine cell populations, including various cancer types and normal cells.
- Cells were exposed to ten clinically used anticancer drugs for 72 hours in microtiter plates.
- Drug potency determined by 50% inhibitory concentration (IC50) based on dye reduction and cell doublings.
Main Results:
- The modified MTT assay demonstrated comparable drug potency (IC50) whether measuring dye reduction or cell doublings.
- High agreement was observed between the MTT assay and the conventional trypan blue exclusion method.
- The assay proved sensitive and reproducible across different cell lines and drug concentrations.
Conclusions:
- The adapted MTT assay is a rapid, sensitive, and reproducible first-line screening tool for anticancer compounds.
- This high-capacity system facilitates efficient detection of compounds with cytostatic or cytocidal activity.
