流式细胞术在小细胞外囊泡检测中的研究进展

Research progress of flow cytometry in detection of small extracellular vesicles

摘要:

小细胞外囊泡（small extracellular vesicles，sEV）是由细胞主动分泌、尺寸小于200 nm、具有脂质双分子膜结构的球状膜性微小囊泡的总称，sEV参与细胞间信号通讯，在生理状态维持及疾病进程中发挥重要作用，是近年来学界研究的热点。流式细胞术具有能直接对大量细胞等进行多参数、快速度、高分辨率分析的优势，但应用在检测比细胞小很多的sEV研究中还较少。本文综述了多个研究团队报道的将流式细胞术应用于检测sEV的方法，主要包括纳米流式细胞术直接检测sEV；通过sEV聚类、DNA扩增、微球等辅助常规流式细胞仪实现检测sEV；质谱流式细胞术辅助检测sEV等。以上方法显示出了流式细胞术应用于sEV检测的可能与优势，同时也面临着如荧光光谱重叠干扰、无法准确定量、依然不能突破尺寸限制等问题，限制了其大规模的研究应用。希望本综述能帮助学界更好地了解sEV流式细胞术的研究进展，进一步推动sEV的研究。

Abstract:

Small extracellular vesicles（sEV） are spherical membranous vesicles with lipid bilayer membrane structure and size less than 200 nm，which are secreted by cells actively. sEV are involved in intercellular signal communication and play an important role in the maintenance of normal physiological state and the progression of disease. Flow cytometry has the advantage of being able to directly perform multi-parameter，fast and high-resolution analysis on a large number of cells，but few studies have been applied to detect sEV that are much smaller than cells. In this paper，we review the application of flow cytometry in the detection of sEV reported by several research teams，including the direct detection of sEV by nanoflow cytometry. By conventional flow cytometry assisted by sEV clustering，DNA amplification and microspheres. By mass spectrometry flow cytometry. These methods show the possibility and advantages of flow cytometry in the detection of sEV，but at the same time，they are also faced with problems such as overlapping interference of fluorescence spectrum，and inability to accurately quantify，and still unable to break through size limits，which limit its large-scale research and application. This review is expected to help the academic community better understand the research progress of sEV flow cytometry and further promote the research of sEV.