Multispectral Optical Tweezers for Biochemical Fingerprinting of CD9-Positive Exosome Subpopulations.
Multispectral Optical Tweezers for Biochemical Fingerprinting of CD9-Positive Exosome Subpopulations.
复制标题
DOI：
10.1021/acs.analchem.7b00017
复制
发表时间：
2017-05-16
影响因子：
7.4
通讯作者：
Lam KS
中科院分区：
文献类型：
--
作者：
Carney RP;Hazari S;Colquhoun M;Tran D;Hwang B;Mulligan MS;Bryers JD;Girda E;Leiserowitz GS;Smith ZJ;Lam KS
细胞外囊泡（EV），包括外来体，是循环的纳米级颗粒，与细胞信号传导密切相关，并且可以从人类生物流体中大量分离。目前正在研究它们的分子特征和材料特性，以描述各种生物功能和疾病。然而，大量的，但在很大程度上尚未量化，各种EV亚群的组成，大小和可能的功能不同，需要能够测量单个囊泡的表征方案。在这里，我们描述的第一个应用程序的多光谱光镊（MS-OTs）的单囊泡的EV亚群的分子指纹。该多功能成像平台允许对拉曼化学成分（例如，蛋白质、脂质、胆固醇、核酸的变化），再加上荧光标记物的区分。对于通过超离心分离的外泌体，我们使用MS-OT通过抗体荧光标记和拉曼光谱测量来询问CD 9阳性亚群。我们报告称，与总纯化EV群体相比，CD 9阳性外泌体子集显示每个囊泡的成分浓度降低，化学异质性降低。我们观察到，特定的囊泡亚群存在于从间充质基质细胞的几种克隆品种的细胞培养上清液中分离的外泌体中，也存在于从人卵巢癌患者分离的血浆和腹水中。
Extracellular vesicles (EVs), including exosomes, are circulating nanoscale particles heavily implicated in cell signaling and can be isolated in vast numbers from human biofluids. Study of their molecular profiling and materials properties is currently underway for purposes of describing a variety of biological functions and diseases. However, the large, and as yet largely unquantified, variety of EV subpopulations differing in composition, size, and likely function necessitates characterization schemes capable of measuring single vesicles. Here we describe the first application of multispectral optical tweezers (MS-OTs) to single vesicles for molecular fingerprinting of EV subpopulations. This versatile imaging platform allows for sensitive measurement of Raman chemical composition (e.g., variation in protein, lipid, cholesterol, nucleic acids), coupled with discrimination by fluorescence markers. For exosomes isolated by ultracentrifugation, we use MS-OTs to interrogate the CD9- positive subpopulations via antibody fluorescence labeling and Raman spectra measurement. We report that the CD9-positive exosome subset exhibits reduced component concentration per vesicle and reduced chemical heterogeneity compared to the total purified EV population. We observed that specific vesicle subpopulations are present across exosomes isolated from cell culture supernatant of several clonal varieties of mesenchymal stromal cells and also from plasma and ascites isolated from human ovarian cancer patients.
登录
查看更多内容
影响因子：
16
作者：
Lötvall J;Hill AF;Hochberg F;Buzás EI;Di Vizio D;Gardiner C;Gho YS;Kurochkin IV;Mathivanan S;Quesenberry P;Sahoo S;Tahara H;Wauben MH;Witwer KW;Théry C
通讯作者：
Théry C
DOI：
10.1073/pnas.1521230113
发表时间：
2016-02-23
影响因子：
11.1
作者：
Kowal, Joanna;Arras, Guillaume;Thery, Clotilde
通讯作者：
Thery, Clotilde
影响因子：
4.6
作者：
Willms E;Johansson HJ;Mäger I;Lee Y;Blomberg KE;Sadik M;Alaarg A;Smith CI;Lehtiö J;El Andaloussi S;Wood MJ;Vader P
通讯作者：
Vader P
影响因子：
4
作者：
Momen-Heravi F;Balaj L;Alian S;Tigges J;Toxavidis V;Ericsson M;Distel RJ;Ivanov AR;Skog J;Kuo WP
通讯作者：
Kuo WP
影响因子：
16
作者：
Lobb RJ;Becker M;Wen SW;Wong CS;Wiegmans AP;Leimgruber A;Möller A
通讯作者：
Möller A