Real-Time Measurement of Mitochondrial Function and Glycolysis in Lymphoblastoid Cell Lines.
Katsaros, Tina, Missailidis, Daniel, Annesley, Sarah J · Methods in molecular biology (Clifton, N.J.) · 2025 · DOI
Quick Summary
This study describes a new method to measure how cells make energy in people with ME/CFS. Researchers use a special machine to track two main energy-production pathways in cells: one that happens quickly (glycolysis) and one that happens in the cell's energy factories called mitochondria. By measuring how much oxygen cells use and the acids they produce, scientists can see if these energy pathways are working differently in ME/CFS patients compared to healthy people.
Why It Matters
Measuring how cells produce energy is crucial for understanding ME/CFS, since abnormal energy metabolism appears to be central to this disease. This standardized method allows researchers worldwide to measure energy dysfunction consistently, potentially leading to better diagnostic tools and new treatments. Having reliable ways to test cell energy function helps scientists identify what goes wrong at the cellular level in ME/CFS.
Observed Findings
The paper provides an optimized Seahorse assay protocol for measuring oxygen consumption rate in lymphoblastoid cell lines
The assay measures extracellular acidification rate to assess glycolytic function and capacity
Sequential injection of metabolic modulators enables measurement of multiple components of both mitochondrial and glycolytic pathways
The protocol has been optimized and validated for use in actively proliferating lymphoblastoid cell lines from blood cells
Inferred Conclusions
Standardized measurement of mitochondrial and glycolytic function in lymphoblastoid cell lines can provide reproducible assessment of cellular bioenergetics in ME/CFS
The use of live, intact cells allows for more physiologically relevant measurement of energy metabolism compared to other approaches
This optimized protocol enables researchers to systematically evaluate different components and stages of energy production pathways
Remaining Questions
How do the bioenergetic profiles measured by this assay correlate with clinical symptoms and disease severity in ME/CFS patients?
Do lymphoblastoid cell lines accurately reflect the energy dysfunction seen in other cell types affected in ME/CFS, such as muscle cells and immune cells?
What This Study Does Not Prove
This methods paper does not provide original research data or clinical findings—it only describes the laboratory technique itself. The study does not prove that energy dysfunction causes ME/CFS symptoms, nor does it establish how cellular energy problems relate to specific patient symptoms. This is a technical protocol paper, not a clinical investigation, so it cannot demonstrate disease mechanisms or treatment efficacy.