Long-term high throughput agitation culturing with real-time metabolic profiling

Abstract

Cellular metabolism relies on the dynamic coordination between glycolytic flux in the cytosol and oxidative phosphorylation (OXPHOS) within the mitochondria. To study the metabolic profiles of cells, researchers apply a monitoring system for measurements of critical parameters, e.g., pH and dissolved oxygen (DO), to understand underlying energy production tendencies, dictating the performance, resilience and growth of cells. However, implementing sensitive, non-invasive sensors into long-term culturing environments remains a technical bottleneck. Here, we describe the DolphinQ bioanalyzer, a novel culturing platform designed for high-throughput, real-time monitoring of cellular metabolism states under physiologically relevant conditions. We validate the system across multiple cell types and experimental set-ups, demonstrating its ability to resolve subtle metabolic shifts that are typically obscured in end-point assays. Notably, we utilize the system to characterize the metabolic impact of heteroplasmy in a mitochondrial disease model with affected ATP synthase. Our results underscore the utility of continuous, minimally disruptive monitoring for revealing the complexities of cellular metabolic adaptation. The DolphinQ framework therefore offers a robust tool for optimizing culture conditions across a wide range of applications and advancing fundamental research into metabolic flux and mitochondrial dysfunction.