Assessment of metabolic activity of human cells in solution and in polymer matrix with the use of metabolite-sensitive sensors

We developed metabolite-sensitive electrochemical sensors on the basis of electrodes modified with a thick film of carbon nanotubes. Modified electrodes provide efficient pre-adsorption of cellular metabolites and their sensitive detection using anodic square-wave voltammetry. On the electrode surface both adhered and non-adhered human cells produce three oxidation peaks at the potentials of + 0.82, + 1.05, and + 1.17 V attributed to three groups of cellular metabolites: amino acid-derived antioxidants including glutathione, guanine nucleotides, and also adenine nucleotides including ATP. The electrochemical response was well correlated with cell viability, intracellular ATP level and induction of apoptosis, as determined by independent assays. Developed sensors allow for robust and cost-effective assessment of ATP in cells in contrast to enzyme-based electrodes and conventional bioluminescent assay. Results can be used for rapid analysis of human cells for the purpose of medical diagnostics, transplantology, and toxicological screening. Additionally, we combined modified electrodes with human cells entrapped in agarose matrix. The resulting biosensor allowed for electrochemical monitoring of metabolic activity and death of cells within polymeric matrix that is of interest for tissue engineering applications.