Characterization of ROCK inhibitor-treated extracellular vesicles and analysis of their effect on osteogenesis

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MagCapture Exosome Isolation Kit was used to prepare Control extracellular vesicles (Control-EVs) and ROCK inhibitor-treated extracellular vesicles (ROCKin'-EVs) from the culture medium of BMSCs. Mass spectrometry was performed to elucidate the changes in the proteomic composition. We detected there were changes ROCKin'-EVs protein composition. We identified that a number of Rab GTPase (known as the key regulators of intercellular vesicle transport) were upregulated in ROCKin'-EVs.We isolated the Control-EVs and ROCKin'-EVs from the culture medium of BMSCs again using total exosome isolation aiming to collect large amount of EVs. Western blotting of Control-EVs and ROCKin'-EVs was performed. The quantity of isolated EVs was inconsistent and necessary to be normalized.