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Abstract
A colorimetric microtiter assay was developed for the quantitation of adherent cells
in culture, which is based on the staining of cells with a commercially available
staining kit. Adherent L929, A375 cell lines and human monocytes were stained with
Hemacolor reagents and the color eluted with SDS 0.5% was determined spectrophotometrically
with an ELISA plate reader at 630 nm. The method enabled the detection of cells and
was linear up to 3 x 10(4) L929 cells/well. The Hemacolor staining assay was compared
to the crystal violet staining assay and the MTT reduction assay, and was found to
be sensitive, accurate and reproducible, and has the advantage of enabling microscopic
inspection of the stained cells prior to color elution. The assay was found to be
suitable for the determination of cytotoxic cytokines, and the enumeration of adherent
monocytes. This method might be also used for the quantitation of cytotoxic drugs,
and the cytophatic activity of viruses.