Antiproliferative drugs and human ocular fibroblasts: colorimetric vs. cell counting assays

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Abstract
Utilizing a rapid colorimetric assay that relates cell number to cytoplasmic hexosaminidase activity, we conducted drug-induced cytotoxicity experiments on human ocular fibroblasts cultured from Tenon's capsule specimens. The effects of two different agents –dimethyl sulfoxide (DMSO) or mitomycin C– on the proliferation of human ocular fibroblasts were studied. Simultaneously the sensitivity of this technique was compared to electronic cell counting with a Coulter counter, a conventional means of quantifying proliferation. Known numbers of cells were exposed to varied concentrations of either DMSO or mitomycin C for 11 days. Cell attachment was quantified after 24 hours, and proliferation was quantified periodically thereafter over the remaining 10 days. Colorimetric data contained a similar or smaller amount of random error than corresponding Coulter values. Both assays identified statistically significant antiproliferative effects and inhibitory effects on cell attachment at higher drug doses; however, Coulter counting alone detected many additional significant effects among lower-dose DMSO and mitomycin treatment groups. Although the hexosaminidase assay displayed lower sensitivity than Coulter counting, it may still be useful to rapidly screen new compounds for strong antifibroblastic effects.