Vella Laura J, Scicluna Benjamin J, Cheng Lesley, Bawden Emma G, Masters Colin L, Ang Ching-Seng, Willamson Nicholas, McLean Catriona, Barnham Kevin J, Hill Andrew F
The Florey Institute of Neuroscience and Mental Health, The University of Melbourne, Parkville, Australia.
The Bio21 Molecular Science and Biotechnology Institute, The University of Melbourne, Parkville, Australia.
J Extracell Vesicles. 2017 Jul 26;6(1):1348885. doi: 10.1080/20013078.2017.1348885. eCollection 2017.
Extracellular vesicles, including exosomes, are released by all cells, including those of the nervous system. Capable of delivering lipid, protein and nucleic acids to both nearby and distal cells, exosomes have been hypothesized to play a role in progression of many diseases of the nervous system. To date, most analyses on the role of these vesicles in the healthy and diseased state have relied on studying vesicles from sources, such as conditioned cell culture media, or body fluids. Here we have taken a critical approach to the enrichment and characterization of exosomes from human frontal cortex. This method maintains the integrity of the vesicles and their cargo, and comprehensive proteomic and genomic characterization confirms the legitimacy of the resulting extracellular vesicles as endosome-derived exosomes. This method will enable neuroscientists to acquire more detailed information about exosomes in the brain and explore the role(s) this form of intercellular communication and unique source of lipid, protein and RNA has in healthy brain function and pathogenic conditions. Furthermore, this method may have important utility in the isolation of exosomes from other tissues.
包括外泌体在内的细胞外囊泡由所有细胞释放，包括神经系统的细胞。外泌体能够将脂质、蛋白质和核酸传递给附近和远处的细胞，据推测其在许多神经系统疾病的进展中发挥作用。迄今为止，大多数关于这些囊泡在健康和疾病状态下作用的分析都依赖于研究来自条件细胞培养基或体液等来源的囊泡。在这里，我们采用了一种关键方法来富集和表征来自人类额叶皮质的外泌体。该方法保持了囊泡及其货物的完整性，全面的蛋白质组学和基因组学表征证实了所得细胞外囊泡作为内体来源外泌体的合法性。这种方法将使神经科学家能够获得有关大脑中外泌体的更详细信息，并探索这种细胞间通讯形式以及独特的脂质、蛋白质和RNA来源在健康脑功能和致病条件中的作用。此外，这种方法在从其他组织中分离外泌体方面可能具有重要用途。
