Pitfalls associated with lipophilic fluorophore staining of extracellular vesicles for uptake studies
Pitfalls associated with lipophilic fluorophore staining of extracellular vesicles for uptake studies
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DOI：
10.1080/20013078.2019.1582237
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发表时间：
2019-01-01
影响因子：
16
通讯作者：
Simonsen, Jens B.
中科院分区：
文献类型：
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作者：
Simonsen, Jens B.
使用 PKH67 等脂质锚定荧光团 (LAF) 对细胞外囊泡 (EV) 进行后染色是研究 EV 的一种广泛使用的策略，但它存在一些缺陷。本评论中讨论的陷阱与非 EV 物种的 LAF 标记有关，原因是 (1) EV 样品中的脂蛋白污染，(2) LAF 报告基因从囊泡解吸到血液和血清中的蛋白质和脂蛋白中，以及 (3) 两亲性 LAF 化合物形成 EV 样颗粒的能力。意识到这些挑战并制定解决方案来克服这些挑战对于确保我们在使用 LAF 跟踪电动汽车时做出相关解释非常重要。
Post-staining of extracellular vesicles (EVs) with lipid-anchored fluorophores (LAFs) such as PKH67 is a widely used strategy for studying EVs but it is associated with several pitfalls. The pitfalls discussed in this commentary are related to LAF labelling of non-EV species due to (1) lipoprotein contamination in EV samples, (2) desorption of the LAF reporters from vesicles into proteins and lipoproteins in blood and serum, and (3) the capability of the amphiphilic LAF compounds to form EV-like particles. Awareness of these challenges and developing solutions to overcome these are important to ensure that we make relevant interpretations when using LAFs to track EVs.