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Updated: Aug 6, 2026
Size Exclusion Chromatography for Separating Extracellular Vesicles from Conditioned Cell Culture Media
Published on: May 13, 2022
Protocol for ultracentrifugation-based isolation and characterization of extracellular vesicles from FGF2-primed
Xiaomin Wen1, Mingming Wang1, Zihao Gao1
1Department of Neurobiology, School of Basic Medicine, Fourth Military Medical University, Xi'an, Shaanxi 710032, P.R. China; The Shaanxi Province Key Laboratory of Brain Function Analysis and Modulation, Xi'an, Shaanxi 710032, P.R. China.
Abstract:
Extracellular vesicles (EVs) mediate intercellular communication by transferring bioactive molecular cargo. We present a protocol for isolating EVs from fibroblast growth factor-2 (FGF2)-primed primary mouse astrocytes using differential ultracentrifugation. We describe steps for primary astrocyte culture, FGF2 priming, EV isolation, and comprehensive characterization encompassing transmission electron microscopy (TEM). We then detail procedures for nanoparticle tracking analysis (NTA), western blotting, and mass spectrometry-based proteomic analysis. EVs isolated by this approach protect against mitochondrial and synaptic toxicity in Parkinson's disease models. For complete details on the use and execution of this protocol, please refer to Wen et al.1.