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

用MagCapture Exosome分离试剂盒从BMSCs的培养液中制备对照胞外囊泡(Control-EVS)和ROCK抑制剂处理的胞外囊泡(Rockin‘-EVS)。用质谱仪分析蛋白质组分的变化。我们检测到Rockin‘-EVS的蛋白质组成发生了变化。我们发现Rockin-EV中存在多种Rab GTP酶(Rab GTP酶是细胞间囊泡运输的关键调节因子)。为了收集大量EV，我们再次从BMSCs的培养液中分离出对照EVS和Rockin‘-EVS。对Control-EVS和Rockin‘-EVS进行蛋白质印迹。分离的EVS数量不一致，有必要进行标准化。

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.