Institute of Pharmacy-Pharmaceutical Biology, Dahlem Center of Plant Sciences, Freie Universitaet Berlin, Koenigin-Luise-Str. 2+4, D-14195 Berlin, Germany.
Int J Mol Sci. 2019 Jan 16;20(2):357. doi: 10.3390/ijms20020357.
Although animal-derived extracellular vesicles (EVs) are moving increasingly into scientific focus, EVs from other kingdoms remain underestimated and our knowledge of them is still expandable, probably due to the lack of an easy and broadly executable isolation, purification and visualization method. Using differential centrifugation with subsequent agarose gel electrophoresis, we were able to simplify the terms of EV isolation. EVs from L., L., and L. were purified, even though they did not migrate into the gel matrix. If 3,3- Dihexyloxacarbocyanine iodide (DiOC 6 ) is added to the specimen in excess, membranous components can already be detected by eye, or with higher sensitivity, using a UV transilluminator. The sample preparation can be adjusted to the EV species of interest. Moreover, EVs are separated from small charged contaminants and dye excess, because these impurities can pass the gel matrix, while EVs themselves are retained in the pocket. Significantly, we isolated EVs from dried plant material, which is-to our knowledge-the first proof that EVs are stable enough to overcome the drying process of plant material.
尽管动物来源的细胞外囊泡 (EVs) 越来越受到科学界的关注，但来自其他生物界的 EVs 仍被低估，我们对它们的了解仍在不断扩展，这可能是由于缺乏简单且广泛可行的分离、纯化和可视化方法。我们使用差速离心和随后的琼脂糖凝胶电泳，成功地简化了 EV 分离的条件。即使没有迁移到凝胶基质中，我们也能够从 L.、L. 和 L. 中纯化出 EV。如果向样本中过量添加 3,3-二己氧基羰花青碘化物 (DiOC6)，可以通过肉眼或使用紫外透射仪以更高的灵敏度检测到膜性成分。样品制备可以根据感兴趣的 EV 种类进行调整。此外，EV 可以与小的带电荷的污染物和染料过剩物分离，因为这些杂质可以通过凝胶基质，而 EV 本身则被保留在口袋中。值得注意的是，我们从干燥的植物材料中分离出 EV，这是我们所知的第一个证明 EV 足够稳定，可以克服植物材料的干燥过程的证据。
