Assessing extracellular vesicles in human biofluids using flow-based analyzers
Assessing extracellular vesicles in human biofluids using flow-based analyzers
复制标题
使用基于流的分析仪评估人体生物液中的细胞外囊泡
DOI：
10.1101/2022.07.20.500853
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发表时间：
2022
期刊：
影响因子：
--
通讯作者：
Krzyzaniak O
中科院分区：
文献类型：
--
作者：
Krzyzaniak O
细胞外囊泡（EVs）越来越多地被用于流式细胞术分析。然而，由于其微小的尺寸和低折射率，导致大多数ev的散射强度低于大多数流式细胞仪的检测极限。一类新的设备，被称为光谱流分析仪，正在成为细胞表型研究的标准。很大程度上，由于其独特的能力，检测一个巨大的面板标记具有更高的灵敏度光散射检测。另一类设备，被称为纳米分析仪，提供亚微米大小的颗粒的高分辨率检测。在这里，我们的目的是比较极光（Cytek）光谱细胞分析仪和纳米ofcm （nFCM）纳米流分析仪之间的ev表型表现。这两种设备被特别选择，因为它们在各自领域成为黄金标准的领先地位。尽管免疫细胞衍生的ev具有临床潜力，但其特征仍然很差。因此，我们使用B和T细胞系衍生的ev和供体匹配的人类生物体液衍生的ev，这些ev来自血清、尿液和唾液，并结合一组已建立的免疫标记物进行比较研究。对两种细胞仪平台进行了比较评估，讨论了它们在不同应用中的潜力和适用性。我们发现nFCM可以准确地分析小型ev (40 ~ 200 nm)，与电子显微镜的尺寸精度相匹配；ii)测量每体积内单个EV颗粒的浓度；iii)识别未被充分代表的EV标记子集；iv)提供EV表面标记的共定位。我们还可以证明人类生物体液样本具有独特的EV标记物特征，这可能具有未来的临床相关性。
Extracellular vesicles (EVs) are increasingly being analyzed by flow cytometry. Yet, their miniscule size and low refractive index, causes the scatter intensity of most EVs to fall below the detection limit of most flow cytometers. A new class of devices, known as spectral flow analyzers, are becoming standards in cell phenotyping studies. Largely, due to their unique capacity of detecting a vast panel of markers with higher sensitivity for light scatter detection. Another class of devices, known as nano-analyzers, provides high resolution detection of sub-micron sized particles. Here, we aim to compare the EVs phenotyping performance between the Aurora (Cytek) spectral cell analyzer and the NanoFCM (nFCM) nanoflow analyzer. These two devices were specifically chosen given their lead in becoming gold standards in their respective fields. Immune cell-derived EVs remain poorly characterized despite their clinical potentials. We therefore, used B- and T- cell line-derived EVs and donor-matched human biofluid-derived EVs from serum, urine, and saliva in combination with a panel of established immune markers for this comparative study. A comparative evaluation of both cytometry platforms was performed, discussing their potential and suitability for different applications. We found that nFCM can accurately i) analyze small EVs (40 to 200 nm) matching the size accuracy of electron microscopy; ii) measure concentration of single EV particle per volume; iii) identify underrepresented EV marker subsets; and iv) provide co-localization of EV surface markers. We could also show that human sample biofluids have unique EV marker signatures that could have future clinical relevance.
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DOI：
10.1002/jex2.37
发表时间：
2022-04
期刊：
Journal of extracellular biology
影响因子：
--
作者：
通讯作者：
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影响因子：
9.3
作者：
de Rond, Leonie;van der Pol, Edwin;Coumans, Frank A. W.
通讯作者：
Coumans, Frank A. W.
影响因子：
5.6
作者：
Lucchetti D;Battaglia A;Ricciardi-Tenore C;Colella F;Perelli L;De Maria R;Scambia G;Sgambato A;Fattorossi A
通讯作者：
Fattorossi A
影响因子：
16
作者：
Silva AM;Lázaro-Ibáñez E;Gunnarsson A;Dhande A;Daaboul G;Peacock B;Osteikoetxea X;Salmond N;Friis KP;Shatnyeva O;Dekker N
通讯作者：
Dekker N
影响因子：
16.6
作者：
Khairnar, Vishal;Duhan, Vikas;Maney, Sathish Kumar;Honke, Nadine;Shaabani, Namir;Pandyra, Aleksandra A.;Seifert, Marc;Pozdeev, Vitaly;Xu, Haifeng C.;Sharma, Piyush;Baldin, Fabian;Marquardsen, Florian;Merches, Katja;Lang, Elisabeth;Kirschning, Carsten;Westendorf, Astrid M.;Hauessinger, Dieter;Lang, Florian;Dittmer, Ulf;Kueppers, Ralf;Recher, Mike;Hardt, Cornelia;Scheffrahn, Inka;Beauchemin, Nicole;Goethert, Joachim R.;Singer, Bernhard B.;Lang, Philipp A.;Lang, Karl S.
通讯作者：
Lang, Karl S.