A Novel Cytotoxicity Screening Assay Using a Multiwell Fluorescence Scanner
| dc.contributor.author | Anna-Liisa Nieminen | |
| dc.contributor.author | Gregory J. Gores | |
| dc.contributor.author | John M. Bond | |
| dc.contributor.author | Roberto Imberti | |
| dc.contributor.author | Brian Herman | |
| dc.contributor.author | John J. Lemasters | |
| dc.date.accessioned | 2014-09-08T08:55:31Z | |
| dc.date.available | 2014-09-08T08:55:31Z | |
| dc.date.issued | 1992 | |
| dc.description.abstract | A new assay using a multiwell fluorescence scanner was developed for screening Cytotoxicity to cells cultured in 96-well microtiter plates. The assay is based on binding of propidium i 'dide to nuclei of cells whose plasma membranes have become permeable due to cell death. Fluorescence of propidium iodide measured with a multiwell fluorescence scanner increased in proportion to the number of permeabilized cells. After ATP depletion of hepatocytes and neonatal cardiac myocytes with metabolic inhibitors ("chemical hypoxia"), and exposure of Madine Darby canine kidney cells to the toxic chemical, HgCI2, propidium iodide fluorescence progressively increased. Increases of fluorescence were linearly proportional with release of lactate oehydrogenase into the culture medium. Employing this cyto-toxicity screening assay, protection by various agents against lethal injury was evaluated in cultured hepatocytes during chemical hypoxia. Inhibitors of cysteine proteases (i.e., antipain, leu-peptin, E-64), serine proteases (i.e., PMSF). and aspartic acid proteases (i.e., pepstatin A) did not protect against chemical hypoxia. In contrast, 1,10-phenanthroline, an inhibitor of me-talloprotease, markedly protected against the onset of cell death during chemical hypoxia. Half-maximal protection after 60 min occurred at 0.5 MM. Phospholipase inhibitors, chlorpromazine (50 MM) and mepacrine (50 MM), also substantially retarded cell killing. U74006F, an inhibitor of lipid peroxidation, slowed cell killing to a lesser extent during chemical hypoxia and after ox-idative stress with f-butyl hydroperoxide. Calciphor, a dimer of p)-ostaglandin B1, did not protect against cell killing during cnemical hypoxia or r-butyl hydroperoxide toxicity. In conclusion, this high capacity cytoloxicity assay for cells cultured in 96-wcll microtiter plates is suitable for rapid screening of potential cyloprotective agents in a variety of cell types. | en_US |
| dc.identifier.citation | Toxicology and Applied Pharmacology 115, 147-155, 1992 | en_US |
| dc.identifier.uri | http://hdl.handle.net/123456789/5479 | |
| dc.language.iso | en | en_US |
| dc.title | A Novel Cytotoxicity Screening Assay Using a Multiwell Fluorescence Scanner | en_US |
| dc.type | Article | en_US |