Pursuing mechanisms of extracellular vesicle formation. Effects of sample processing
Pursuing mechanisms of extracellular vesicle formation. Effects of sample processing
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DOI：
10.1016/bs.abl.2020.09.003
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发表时间：
2020-01-01
期刊：
影响因子：
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通讯作者：
Kralj-Iglic, Veronika
中科院分区：
文献类型：
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作者：
Bozic, Darja;Hocevar, Matej;Kralj-Iglic, Veronika
尽管细胞外囊泡在细胞间通讯中的作用已得到公认，但它们的分析仍存在问题。对样本处理效果的了解不足可能是这个问题的基石之一。我们研究了在囊泡分离过程中离心对分离物中颗粒的数量、大小、含量和形状的影响。通过扫描电子显微镜观察，在约100-500 nm范围内的颗粒表现出没有内部结构的囊泡的特征形状。离心力的增强导致分离物中某些类型蛋白质的量增加。大多数蛋白质是高丰度的人血浆蛋白，包括白蛋白和免疫球蛋白。结果是在支持的假设，即孤立的颗粒反映了他们来自（细胞和周围的解决方案）和执行的处理的物质，因此，囊泡在隔离物应被视为一个动态的材料与瞬态身份。
Although the role of extracellular vesicles in inter-cell communication is acknowledged, their analytics is yet problematic. Poor understanding of effects of sample processing may be one of the cornerstones of this issue. We investigated the effects of centrifugation during the vesicle isolation on the number, size, content and shape of particles in the isolates. Particles in the range of about 100-500 nm as observed by the scanning electron microscope exhibited characteristic shapes of vesicles without internal structure. Enhanced centrifugal pull resulted in increased amount of certain types of proteins in the isolates. Majority of present proteins were highly abundant human plasma proteins, including albumins and immunoglobulins. Results are in favor of the hypothesis that the isolated particles reflect the substance from which they originate (cells and the surrounding solution) and the performed processing; and therefore, vesicles in isolates should be viewed upon as a dynamic material with transient identity.