Isolation and Analysis of Traceable and Functionalized Extracellular Vesicles from the Plasma and Solid Tissues

Area of Science:

• Biochemistry
• Cell Biology
• Biomarker Discovery

Background:

• Extracellular vesicles (EVs) are crucial in homeostasis and disease, with both exosomes and microvesicles gaining research attention.
• Microvesicles, originating from plasma membrane blebbing, reflect the molecular signature of parent cells.
• Current research often emphasizes exosomes, necessitating better characterization of other EV types.

Purpose of the Study:

• To present a reproducible protocol for the isolation and characterization of extracellular vesicles (EVs) from biological fluids and solid tissues.
• To enable the profiling of surface antigens and protein cargo within EVs for functional analysis.
• To facilitate the traceability of EVs to their cellular origins and understand their roles in physiological and pathological processes.

Main Methods:

• Differential centrifugation for the extraction of EVs from plasma and solid tissues (e.g., bone).
• Characterization of isolated EVs, including profiling of surface markers and protein content.
• Methodology designed for reproducibility and broad applicability in biological studies.

Main Results:

• A reproducible procedure for isolating and characterizing EVs from diverse sources, including plasma and bone tissue.
• The protocol allows for detailed profiling of EV surface molecules and protein cargo.
• Enables tracing EV origins and identifying functional components.

Conclusions:

• The presented method provides a robust approach for EV isolation and characterization.
• This protocol supports correlative, functional, and mechanistic studies of EVs in various biological contexts.
• Facilitates the use of EVs as theranostic biomarkers in health and disease research.