Method to Determine Quality of Mitochondrial Cargo in Extracellular Vesicles

Background:

Studies have shown that EV-mediated intercellular communication is influenced, at least in part, by the mitochondrial cargo contained within EVs. This highlights the importance of rigorous characterization of both the load and quality of mitochondrial cargo. However, conventional methods such as transmission electron microscopy and proteomic analysis can compromise mitochondrial integrity, limiting the ability to assess cargo functionally. Although single-particle flow cytometry offers promise for mitochondrial cargo characterization, standardized protocols and technical guidance remain insufficient.

Technology Overview:

Researchers at the University at Buffalo have developed a standardized workflow for assessing the quantity and functional quality of mitochondrial cargo within extracellular vesicles (EV) at the single-vesicle level. This protocol identifies intact EV, distinguishes the mitoEV subset (EV carrying mitochondrial cargo), and assesses whether the mitochondrial cargo is functionally polarized or depolarized, reflecting its quality and respiratory competence.

Advantages:

• Workflow is reproducible
• Adaptable to different EV sources
• Provides functional insight unavailable from traditional EV characterization methods (e.g. transmission electron microscopy, western blot, nanoparticle tracking analysis)

Applications:

• Research kits and assays
• Clinical assays
• Therapeutic quality control tools
• Drug development screens
• Fee-for-service testing

Stage of Development:

TRL 4

Licensing Status: 

Available for license or collaboration.