Extracellular Vesicle Isolation and Characterization for Applications in Cartilage Tissue Engineering and...

Extracellular vesicles (EVs) have the capacity for use in cartilage tissue engineering by stimulating tissue repair and microenvironmental reprogramming. This makes them ideal candidates for treating focal cartilage defects and cartilage degeneration in osteoarthritis (OA). Observational studies have reported beneficial biological effects of EVs, such as inhibition of inflammation, enhanced extracellular matrix deposition, and reduced cartilage degradation. Isolation of EVs derived from different source materials such as conditioned cell culture media or biofluids is essential to attribute observed biological effects to EVs as genuine effectors. This chapter presents a density- and a size-based method as well as a combination of both for isolation of EVs from conditioned cell culture media or biofluids. In addition, three methods for characterization of isolated EVs are suggested based on physical properties, protein profiling, and ultrastructural morphology.