Title: Online Monitoring of BALB/3T3 Metabolism and Adhesion with Multiparametric Chip-Based System
Citation: ANALYTICAL BIOCHEMISTRY vol. 371 no. 1 p. 92-104
Publication Year: 2007
JRC Publication N°: JRC42241
ISSN: 0003-2697
URI: http://publications.jrc.ec.europa.eu/repository/handle/JRC42241
DOI: 10.1016/j.ab.2007.07.014
Type: Articles in Journals
Abstract: A multiparametric chip-based system was employed to measure cell adhesion, metabolism, and response to metal compounds previously classified as cytotoxic in immortalized mouse fibroblasts (BALB/3T3 cell line). The system measures in parallel, online, and in label-free conditions the extracellular acidification rates (with pH-sensitive field effect transistors [ISFETs]), the cellular oxygen consumption (with amperometric electrode structures [Clark-type sensors]), and cell adhesion (with impedimetric interdigitated electrode structures [IDESs]). The experimental protocol was optimized to monitor metabolism and adhesion of the BALB/3T3 cell line. A total of 70,000 cells and a bicarbonate buffer-free running low-glucose Dulbecco's modified Eagle's medium (DMEM) containing 10% fetal clone serum III and 1 mM Hepes were selected to maintain cells in good conditions on the chip during the measurements performed under perfusion conditions. Cells were exposed to sodium arsenite, cadmium chloride, and cis-platinum at concentrations ranging from 1 to 100 lM. The kinetics of cell response to these compounds was analyzed and suggests that the Clark-type sensors can be more sensitive than IDESs and ISFETs in detecting the presence of high chemical concentration when short exposure times (i.e., 2 h) are considered. The cytotoxicity data obtained from the online measurements of acidification, respiration, and adhesion at 24 h compare well, in terms of half-inhibition concentration values (IC50), with the ones obtained using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) test and colony-forming efficiency (CFE) assay. The results show a good sensitivity of the system combined with the advantages of the online and label-free detection methods that allow following cell status before, during, and after the treatment in the same experiment.
JRC Institute:Institute for Health and Consumer Protection

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