Immobilisation of Neuro-2a cells on electrodes and electrochemical detection of MTT formazan crystals to assess their viability
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Marine toxins are potent toxic compounds that may reach humans and poison them. Therefore, their detection in seafood is crucial to prevent intoxication cases. Colorimetric cell-based assays (CBAs) have been developed to analyse marine neurotoxins, such as ciguatoxins (CTXs) and tetrodotoxins (TTXs), and are based on the toxicological effect of these toxins on the cells. Cell viability can be quantified by measuring the mitochondrial activity with 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT). With the purpose of moving forward in the development of cell-based biosensors (CBBs) for neurotoxins, Neuro-2a cells were immobilised on electrodes of different materials (carbon, carbon/polyaniline, carbon/poly-L-lysine, carbon/poly(3,4-ethylenedioxythiophene) and gold) and their presence and viability were assessed by the detection of MTT formazan crystals with cyclic voltammetry. Best results in terms of oxidation potential and current intensity were achieved with carbon and carbon/polyaniline electrodes. Light microscopy also proved the presence of immobilised and living cells on electrodes. Cell density, incubation time and MTT concentration were optimised. Appropriate electrochemical responses were obtained incubating 100,000 cells/electrode for 2 h and using 0.86 mg/mL MTT. The system was able to detect toxicity when exposed to CTX1B and TTX standard solutions as well as Seriola dumerili and Lagocephalus sceleratus fish extracts containing these toxins.
574 - Ecologia general i biodiversitat
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Alkassar, Mounira, Sandra Leonardo, Jorge Diogène, and Mònica Campàs. 2022. "Immobilisation Of Neuro-2A Cells On Electrodes And Electrochemical Detection Of MTT Formazan Crystals To Assess Their Viability". Bioelectrochemistry, 108274. doi:10.1016/j.bioelechem.2022.108274.
Grant agreement number
MICINN/Programas Estatales de generación de conocimiento y fortalecimiento científico y tecnológico del sistema de I+D+i y de I+D+i orientada a los retos de la sociedad, convocatoria 2020/PID2020-112976RB-C21/ES/Biotechnological tools based on cells and aptamers for the detection of ciguatoxins and tetrodotoxins/CELLECTRA
MICIU/Programa Estatal de generación del conocimiento y fortalecimiento científico y tecnológico del sistema I+D+I/PID2019-108781RR-C21/ES/Los riesgos alimentarios asociados con las ciguatoxinas: seguimiento y supervisión de las toxinas marinas y organismos productores de toxinas en los ecosistemas marinos/CIGUARISK
Aigües Marines i Continentals
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