Flow cytometric methods used as screening tests for basal toxicity of chemicals

Abstract

Aim of the present study was to evaluate the suitability of flow cytometry to test in vitro effects of toxicants. Flow cytometry offers the possibility to study several parameters simultaneously, e.g. cell cycle modulation, apoptosis and necrosis within the same cell culture. The effects of six compounds (acetaminophen=AAP, isoniazid=INH, digoxin, malathion, paraquat and 2,4-dichlorophenoxy acetic acid=2,4-D) on cell cycle were investigated in HepG2 cells and the induction of apoptosis/necrosis was analyzed by a spectrum of flow cytometric assays in HepG2, AAH-1 and YAC-1 cells. Early indicators of apoptosis––loss of mitochondrial membrane polarization––as well as later events of the apoptotic process––annexin V binding and DNA fragmentation––were studied. The phases of the cell cycle and the occurrence of a sub-G 0 peak of apoptotic cells were determined with propidium iodide staining. The present investigation demonstrated good correlations between results obtained by flow cytometric analyses and the IC50 data of the MEIC (=Multicenter Evaluation of In Vitro Cytotoxicity) study. Regarding the short time required for the tests, the possibility of investigating several parameters of cytotoxicity simultaneously and the ease of performance, flow cytometric analyses are well suited for the pre-screening for toxic effects of chemicals.