Non‐Contact Microfluidic Analysis of the Stiffness of Single Large Extracellular Vesicles from IDH1‐Mutated Glioblastoma Cells

Abstract In preparation for leveraging extracellular vesicles (EVs) for disease diagnostics and therapeutics, fundamental research is being done to understand EV biological, chemical, and physical properties. Most published studies have investigated nanoscale EVs and focused on EV biochemical content. There is much less understanding of large microscale EV characteristics and EV mechanical properties. Recently a non‐contact microfluidic technique that measures the stiffness of large EVs (>1 µm diameter) is introduced. This pilot study probes the robustness of the microfluidic technique to distinguish between EV populations by comparing stiffness distributions of large EVs derived from glioblastoma cell lines. EVs derived from cells expressing the IDH1 mutation, a common glioblastoma mutation known to disrupt lipid metabolism, are stiffer than those expressed from wild‐type cells in a statistical comparison of sample medians. A supporting lipidomics analysis showed that the IDH1 mutation increased the number of saturated lipids in EVs. Taken together, these data encourage further investigation into the potential of high‐throughput microfluidics to distinguish between large EV populations that differ in biomolecular composition. These findings contribute to the understanding of EV biomechanics, in particular for the less studied microscale EVs.