Issue 9, 2025

The impact of common redox mediators on cellular health: a comprehensive study

Abstract

Electrochemistry has become a key technique for studying biomolecular reactions and dynamics of living systems by using electron-transfer reactions to probe the complex interactions between biological redox molecules and their surrounding environments. To enable such measurements, redox mediators such as ferro/ferricyanide, ferrocene methanol, and tris(bipyridine) ruthenium(II) chloride are used. However, the impact of these exogeneous redox mediators on the health of cell cultures remains underexplored. Herein, we present the effects of three common redox mediators on the health of four of the most commonly used cell lines (Panc1, HeLa, U2OS, and MDA-MB-231) in biological studies. Cell health was assessed using three independent parameters: reactive oxygen species quantification by fluorescence flow cytometry, cell migration through scratch assays, and cell growth via luminescence assays. We show that as the concentration of mediator exceeds 1 mM, ROS increases in all cell types while cell viability plumets. In contrast, cell migration was only hindered at the highest concentration of each mediator. Our observations highlight the crucial role that optimized mediator concentrations play in ensuring accuracy when studying biological systems by electrochemical methods. As such, these findings provide a critical reference for selecting redox mediator concentrations for bioanalytical studies on live cells.

Graphical abstract: The impact of common redox mediators on cellular health: a comprehensive study

Supplementary files

Article information

Article type
Paper
Submitted
06 Jan 2025
Accepted
12 Mar 2025
First published
12 Mar 2025
This article is Open Access
Creative Commons BY license

Analyst, 2025,150, 1795-1806

The impact of common redox mediators on cellular health: a comprehensive study

S. P. Nortz, V. Gupta and J. E. Dick, Analyst, 2025, 150, 1795 DOI: 10.1039/D5AN00017C

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