A cell-based biosensor for nanomaterials cytotoxicity assessment in three dimensional cell culture.
Nanoparticles (NPs) are widely used in consumer and medicinal products. The high prevalence of nanoparticles in the environment raises concerns regarding their effects on human health, but there is limited knowledge about how NPs interact with cells or tissues. Because the European Union has called for a substantial reduction of animal experiments for scientific purposes (Directive 2010/63), increased efforts are required to develop in vitro models to evaluate potentially hazardous agents. Here, we describe a new cell-based biosensor for the evaluation of NPs cytotoxicity. The new biosensor is based on transgenic human hepatoblastoma cells (HepG2) that express a secreted form of alkaline phosphatase (SEAP) as a reporter protein whose expression is induced upon activation of a stress response pathway controlled by the transcription regulator nuclear factor-κB (NF-κB). The NF-κB_HepG2 sensor cells were cultured in a Matrigel-based three dimensional environment to simulate the in vivo situation. The new biosensor cells offer the advantage of generating fast and reproducible readout at lower concentrations and shorter incubation time than conventional viability assays, avoid possible interaction between nanomaterials and assay compounds, therefore, minimize generation of false positive or negative results and indicate mechanism of toxicity through NF-κB signaling.
['Dubiak-Szepietowska|Monika|M|', 'Karczmarczyk|Aleksandra|A|', 'Winckler|Thomas|T|', 'Feller|Karl-Heinz|KH|']
D000469:Alkaline Phosphatase D001681:Biological Assay D015374:Biosensing Techniques D018929:Cell Culture Techniques D002470:Cell Survival D056945:Hep G2 Cells D006801:Humans D016328:NF-kappa B D053758:Nanoparticles D020411:Oligonucleotide Array Sequence Analysis D015398:Signal Transduction D018675:Toxicity Tests D014162:Transfection
Cell-based biosensor Nanoparticles NF-κB signaling 3D cell culture