Mediated amperometric monitoring of redox changes during adipocyte differentiation

Real-time monitoring of intracellular redox dynamics provides valuable insight into biological processes such as adipocyte differentiation or adipogenesis. However, assessment of these dynamic metabolic changes in living cells remains technically challenging. Here, we evaluated a DCIP-mediated amperometric sensing for monitoring intracellular redox homeostasis during adipogenesis. Human immortalized and primary adipogenic progenitors were cultured directly on the surface of carbon electrodes. Cells were maintained under adipogenic conditions for 10 days. Cell viability, density, and differentiation were assessed by imaging and cytotoxicity assays at days 0 and 10. Intracellular redox activity was monitored by chronoamperometry following electrode polarization and addition of 2,6-dichlorophenolindophenol (DCIP). DCIP at 15 μM generated a rapid and reproducible amperometric response without detectable cytotoxicity for 30 min. The current stabilized within one minute, increased proportionally with the number of adherent cells and responded to metabolic perturbations demonstrating that the signal reflected cellular metabolic activity. Most importantly, differentiated adipogenic progenitors exhibited higher currents than undifferentiated cells, reflecting associated changes of intracellular electron-transfer activity. Collectively, the findings demonstrate that this DCIP-mediated amperometric platform enables rapid, reproducible and standardized monitoring of intracellular redox changes associated with adipogenesis providing a fluorescent probe-free, sensitive, non-destructive approach for real-time investigation of cellular metabolism and differentiation.