Technical Advances to Study Extracellular Vesicles.
Technical Advances to Study Extracellular Vesicles.
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DOI：
10.3389/fmolb.2017.00079
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发表时间：
2017
影响因子：
5
通讯作者：
O'Loghlen A
中科院分区：
文献类型：
--
作者：
Carpintero-Fernández P;Fafián-Labora J;O'Loghlen A
细胞外囊泡是脂质双层膜纳米颗粒的异质和动态组，其可根据其细胞来源分为三个不同的组：外泌体、微囊泡和凋亡小体。它们由不同的细胞类型产生，可以从几乎所有的体液中分离出来。EV含有多种蛋白质、脂质、核酸和代谢物，其局部和全身调节许多生物和病理情况。已经描述了不同的技术，以确定EV隔离，释放，摄取和货物。尽管免疫印迹、荧光显微镜和电子显微镜等标准技术仍被用于表征和可视化EV，但在过去几年中，出现了更精细的技术。例如，EV摄取可以使用Cre报告方法在单细胞水平特异性地确定，并且已经采用基于生物发光的方法报告来确定EV释放和摄取。此外，这些年来，货物识别技术也有了很大的发展。经典的质谱和下一代测序在过去已经被使用，但现在，这些工具的进步已经促进了EV内容的更深入的表征。在这篇综述中，我们的目标是评估在不同的生物系统中研究EV生物学的标准和最新技术进展。
Extracellular vesicles are a heterogeneous and dynamic group of lipid bilayer membrane nanoparticles that can be classified into three different groups depending on their cellular origin: exosomes, microvesicles, and apoptotic bodies. They are produced by different cell types and can be isolated from almost all body fluids. EVs contain a variety of proteins, lipids, nucleic acids, and metabolites which regulate a number of biological and pathological scenarios both locally and systemically. Different techniques have been described in order to determine EV isolation, release, uptake, and cargo. Although standard techniques such as immunoblotting, fluorescent microscopy, and electron microscopy are still being used to characterize and visualize EVs, in the last years, more fine-tuned techniques are emerging. For example, EV uptake can be specifically determined at a single cell level using the Cre reporter methodology and bioluminescence based-methods reports have been employed to determine both EV release and uptake. In addition, techniques for cargo identification have also enormously evolved during these years. Classical mass spectrometry and next generation sequencing have been used in the past, but nowadays, advances in these tools have facilitated a more in depth characterization of the EV content. In this review, we aim to assess the standard and latest technical advances for studying EV biology in different biological systems.
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DOI：
10.1002/stem.2669
发表时间：
2017-10
期刊：
Stem cells (Dayton, Ohio)
影响因子：
--
作者：
Lee WH;Chen WY;Shao NY;Xiao D;Qin X;Baker N;Bae HR;Wei TT;Wang Y;Shukla P;Wu H;Kodo K;Ong SG;Wu JC
通讯作者：
Wu JC
影响因子：
14.8
作者：
Iraci N;Gaude E;Leonardi T;Costa ASH;Cossetti C;Peruzzotti-Jametti L;Bernstock JD;Saini HK;Gelati M;Vescovi AL;Bastos C;Faria N;Occhipinti LG;Enright AJ;Frezza C;Pluchino S
通讯作者：
Pluchino S
影响因子：
4
作者：
Colombo, Marina;Moita, Catarina;Raposo, Graca
通讯作者：
Raposo, Graca
影响因子：
16
作者：
Haraszti RA;Didiot MC;Sapp E;Leszyk J;Shaffer SA;Rockwell HE;Gao F;Narain NR;DiFiglia M;Kiebish MA;Aronin N;Khvorova A
通讯作者：
Khvorova A
影响因子：
16.6
作者：
Lai, Charles P.;Kim, Edward Y.;Badr, Christian E.;Weissleder, Ralph;Mempel, Thorsten R.;Tannous, Bakhos A.;Breakefield, Xandra O.
通讯作者：
Breakefield, Xandra O.