Isolation and Proteomic Analysis of Microvesicles and Exosomes from HT22 Cells and Primary Neurons.

Exosomes and microvesicles are extracellular vesicles (EVs) released by most cell types. The role of EVs as a method of intercellular communication has led to these vesicles becoming a major area of interest in a variety of scientific fields including neuroscience. Emerging evidence is now demonstrating that the biomolecular composition of EVs, especially exosomes, can play a role in the progression of disease including various neurodegenerative diseases and cancer. In addition to the miRNA profiles of EVs, these vesicles also show interesting changes in protein expression profiles under different physiological and pathological conditions. Characterization of these profiles could prove valuable for both understanding disease pathogenesis and for the discovery of new biomarkers of disease. In this chapter, we describe a protocol for isolation of exosomes and microvesicles from immortalized HT22 cells and primary cortical neurons with sufficient yield and low serum contamination required for downstream analysis and label-free relative quantitation by mass spectrometry.