Comparative analysis of eight cytotoxicity assays evaluated within the ACuteTox Project

Abstract

A comparative analysis of eight cytotoxicity assays [the 3T3 and normal human keratinocytes Neutral Red Uptake (NRU) assay, the primary rat hepatocytes, human HepG2 and 3T3 MTT assay, and the human A.704, SH-SY5Y and HepG2 cells propidium iodide (PI) assay] included in several work packages of the EU Integrated Project ACuteTox, has been carried out. The aim was to evaluate whether cells originating from liver, kidney and brain provided different in vitro acute toxicity results, and the influence of primary liver cells versus cell lines originated from the same tissue. Spearman rank correlation analysis and Hierarchical Cluster Analysis were performed based on the IC50 (50% inhibitory concentrations for the endpoint measured) values generated for 57 chemicals. A relatively large number of neurotoxicants and hepatotoxicants were included which allowed to examine the impact of chemicals with specific tissue toxicity on the results. Our analyses confirmed the similarity between the NRU assays and between the two hepatic cell systems related MTT assays. The type of assay appears to have the greatest influence upon the clustering result regardless of the origin of the cells used. The information provided by the NRU and MTT assays differed from that provided by the PI assay. This approach did not allow to show tissue specific toxicity but it does reveal the effectiveness of the clustering methodology for choosing assays for a testing program for predicting e.g. acute oral toxicity in humans.