Methods for extracellular vesicle isolation from cancer cells.
Methods for extracellular vesicle isolation from cancer cells.
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DOI：
10.20517/cdr.2019.118
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发表时间：
2020
期刊：
影响因子：
--
通讯作者：
Languino LR
中科院分区：
文献类型：
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作者：
Salem I;Naranjo NM;Singh A;DeRita R;Krishn SR;Sirman LS;Quaglia F;Duffy A;Bowler N;Sayeed A;Languino LR
已知细胞释放不同类型的囊泡，如小细胞外囊泡（sev）和大细胞外囊泡（lev）。sev和lev在细胞间通讯、转移前生态位形成和疾病进展中发挥重要作用；两者均可检测细胞培养液和生物液体。sev和lev含有多种蛋白质和RNA货物，据信它们通过内化或与细胞表面蛋白结合影响受体细胞的许多生物学功能。最近已经确定，标准的分离技术，如差示超离心，产生混合的ev群体。然而，据报道，密度梯度超离心允许分离无细胞碎片的sev。本文介绍了从细胞培养基、小鼠和人血浆中分离sev的最常用方法，以及从组织中分离sev的新技术。本文还提供了隔离lev的详细步骤。
Cells are known to release different types of vesicles such as small extracellular vesicles (sEVs) and large extracellular vesicles (LEVs). sEVs and LEVs play important roles in intercellular communication, pre-metastatic niche formation, and disease progression; both can be detected cell culture media and biological fluids. sEVs and LEVs contain a variety of protein and RNA cargo, and they are believed to impact many biological functions of the recipient cells upon their internalization or binding to cell surface proteins. It has recently been established that standard isolation techniques, such as differential ultracentrifugation, yield a mixed population of EVs. However, density gradient ultracentrifugation has been reported to allow the isolation of sEVs without cellular debris. Here, we describe the most common methods used to isolate sEVs from cell culture medium, mouse and human plasma, and a new technique for isolating sEVs from tissues as well. This article also provides detailed procedures to isolate LEVs.