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Alternatives to Laboratory Animals - ATLA

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Comparison of Impedance-based Sensors for Cell Adhesion Monitoring and In Vitro Methods for Detecting Cytotoxicity Induced by Chemicals

Jessica Ponti, Laura Ceriotti, Barbara Munaro, Massimo Farina, Alessio Munari, Maurice Whelan, Pascal Colpo, Enrico Sabbioni and François Rossi

The recent European Commission REACH (Registration, Evaluation and Authorisation of Chemicals) policy outlines a plan for toxicological testing by using alternative non-animal in vitro methods. In this context, there is a need to develop and standardise high-throughput screening (HTS) methods for studying the cytotoxicity induced by chemicals. Electrochemical impedance spectroscopy (EIS) can be considered as a complementary technique to alternative in vitro testing for studying cell adhesion to the substrate, and can give real-time and kinetic information on cell responses to a toxicant. This paper describes the development of a home-made chip based on impedance spectroscopy, and its application in studying the kinetics of BALB/3T3 cell adhesion and the cellular responses to a toxic product as a function of time. Concentrations of sodium arsenite, ranging from 10µM up to 1000µM, were tested in the system, and the results were compared with those obtained with standard protocols used to study basal cytotoxicity induced by chemicals in the BALB/3T3 cell line. The results show that the sensitivity of the developed chip was better than that with the MTT test, with the additional advantages of online monitoring.