Quantitative and qualitative flow cytometric analysis of nanosized cell-derived membrane vesicles

被引：199

作者：

Nolte-'t Hoen, Esther N. M. ^{[1
]}

van der Vlist, Els J. ^{[1
]}

Aalberts, Marian ^{[1
]}

Mertens, Hendrik C. H. ^{[1
]}

Bosch, Berend Jan ^{[2
]}

Bartelink, Willem ^{[2
]}

Mastrobattista, Enrico ^{[3
]}

van Gaal, Ethlinn V. B. ^{[3
]}

Stoorvogel, Willem ^{[1
]}

Arkesteijn, Ger J. A. ^{[4
]}

Wauben, Marca H. M. ^{[1
]}

机构：

[1] Univ Utrecht, Fac Vet Med, Dept Biochem & Cell Biol, NL-3584 CM Utrecht, Netherlands

[2] Univ Utrecht, Fac Vet Med, Dept Virol, NL-3584 CM Utrecht, Netherlands

[3] Univ Utrecht, UIPS, Dept Pharmaceut, NL-3584 CM Utrecht, Netherlands

[4] Univ Utrecht, Fac Vet Med, Dept Immunol, NL-3584 CM Utrecht, Netherlands

来源：

NANOMEDICINE-NANOTECHNOLOGY BIOLOGY AND MEDICINE | 2012年 / 8卷 / 05期

关键词：

Cellular membrane vesicles; Exosomes; Nanovesicles; Microvesicles; Flow cytometry; Dendritic cells; Biomarkers; DENDRITIC CELLS; IMMUNE-RESPONSES; LIGHT-SCATTERING; EXOSOMES; MICROPARTICLES; ANTIGEN; PROTEIN; CANCER; NANOPARTICLES; PARTICLES;

D O I：

10.1016/j.nano.2011.09.006

中图分类号：

TB3 [工程材料学];

学科分类号：

0805 ; 080502 ;

摘要：

Nanosized cell-derived membrane vesicles are increasingly recognized as therapeutic vehicles and high-potential biomarkers for several diseases. Currently available methods allow bulk analysis of vesicles but are not suited for accurate quantification and fail to reveal phenotypic heterogeneity in membrane vesicle populations. For such analyses, single vesicle-based, multiparameter, high-throughput methods are needed. We developed a fluorescence-based, high-resolution flow cytometric method for quantitative and qualitative analysis of nanosized membrane vesicles. Proof of principle was obtained by single-particle analysis of virions and liposomes. Further validation was obtained by quantification of cell-derived nanosized membrane vesicles from cell cultures and body fluids. An important aspect was that the technology was extended to detect specific proteins on individual vesicles. This allowed identification of exosome subsets and phenotyping of individual exosomes produced by dendritic cells (DCs) undergoing different modes of activation. The described technology allows quantitative, multiparameter, and high-throughput analysis of a wide variety of nanosized particles and has broad applications.

引用

页码：712 / 720

页数：9

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