The use of flow cytometric methods in acute and long-term in vitro testing

One principal demand for in vitro screening for toxic effects is the ease of performance and the high throughput of test methods. Flow cytometry offers the possibility to study several parameters simultaneously, e.g. cell cycle modulation, mode of cell death, activity of mitochondria. Aim of the present study was to assess the suitability of flow cytometry for the determination of cytotoxicity of test chemicals. Six chemicals chosen from the MEIC list (acetaminophen, isoniazid, paraquat, malathion, digoxin and 2,4-dichlorophenoxy acetic acid) were tested in HepG2, AAH-1, YAC-1 cells and human lymphocytes. Chemicals were applied for 24, 48 h or 28 days. The phases of the cell cycle were determined and the induction of apoptosis and necrosis was demonstrated by annexin binding, analysis of mitochondrial membrane potential and DNA strand breaks. The results of the present study show that flow cytometric methods are well suited to screen for the cytotoxicity of chemicals, both in adherent cells and cells grown in suspension.