In vitro long-term cytotoxicity testing of 27 MEIC chemicals on Hep G2 cells and comparison with acute human toxicity data

• Published in: Toxicology in vitro Vol. 15; no. 2; pp. 153 - 161
• Main Authors: Scheers, E.M, Ekwall, Ba, Dierickx, P.J
• Format: Journal Article
• Language: English
• Published: Oxford Elsevier Ltd 01-04-2001; Elsevier Science
• Subjects: Alternatives; Biological and medical sciences; Carcinoma, Hepatocellular; Chemical and industrial products toxicology. Toxic occupational diseases; Comparison to human toxicity data; EDIT; General aspects; Hep G2; Humans; Linear Models; Long-term cytotoxicity in vitro; Medical sciences; Predictive Value of Tests; Regression Analysis; Toxicity Tests; Toxicology; Tumor Cells, Cultured; Comparison to human toxicity data; DMEM; RDT; Long-term cytotoxicity in vitro; MEIC; Hep G2; FCS; EDIT; Alternatives; Human; Toxicity; Acute; Chemical compound; Prediction; Cytotoxicity; In vitro; Long term; Dose activity relation; In vivo; Lethal dose 50; Cell line; Hepatocyte; Alternative method; Tumor cell
• ISSN: 0887-2333; 1879-3177
• DOI: 10.1016/S0887-2333(00)00062-X

Within the framework of the EDIT (Evaluation guided Development of In vitro Toxicity and toxicokinetic tests) programme, the long-term cytotoxicity of 27 chemicals was investigated on Hep G2 cells. The first step in the experiments was to determine the PI50
24h of the chemicals. This is the concentration of compound needed to reduce the total protein content by 50% after 24 h of treatment. In the long-term experiments the chemicals were tested in six different concentrations, using the PI50
24h as maximum concentration. The cells were treated twice a week with the same concentration of test compound and were trypsinised and counted once a week (dynamic culture). The number of cells was compared to the number of cells of the control. Three major long-term cytotoxicity patterns could be distinguished. After 6 weeks, the EC50
6ws were determined. This is the concentration of compound needed to reduce the number of cells by 50% after 6 weeks of treatment. These values were compared with the PI50
24h. A good correlation was found for the 27 chemicals (
r
2=0.860). After 6 weeks, the concentration of test compound needed to reduce the total cell protein content by 50% after 24 h after 6 weeks of pretreatment of the cells with a particular concentration of test compound was measured: the PI50
24h-6w. For the majority of compounds there is no difference between the PI50
24h and the PI50
24h-6w. For ethanol, arsenic (III) oxide, verapamil hydrochloride and orphenadrine, the PI50
24h-6w increased in comparison to the PI50
24h. For some compounds a doseresponse was observed, indicating that the cells have become more resistant or more sensitive. Linear regression analysis revealed a good correlation (
r
2=0.709) between the EC50
6w and the human acute toxicity. All these data indicate that a good alternative test may be found for predicting the long-term human toxicity.