IMPROVED SENSITIVITY AND DECREASED SAMPLE-SIZE IN A CYTOTOXICITY TESTFOR BIOMATERIALS - A MODIFIED COLONY MICROASSAY USING A MICROPLATE AND CRYSTAL VIOLET STAINING

Modified Eagle's minimum essential medium supplemented with 5% fetal c alf serum was highly sensitive to cytotoxicity and formed large contro l colonies in the V79 colony assay. A highly sensitive cytotoxicity te st was developed using 96-well microtiter plates. Test chemicals or ex tracts of polyurethane materials containing the same chemicals mere ad ded 24 h after inoculation of cell suspensions. The cells mere fixed a nd stained with crystal violet after additional culture for 6 days (V7 9 cells) or 10 days (Balb/3T3 cells). In terms of sensitivity and rapi d quantitative measurement, this modified colony microassay, using a l ow cell density in 96-well microplates, was superior to various cytoto xicity tests such as colony, growth inhibition, cytolethality, and aga r diffusion assays. (C) 1994 John Wiley & Sons, Inc.