TY - JOUR
T1 - Quantitative and qualitative flow cytometric analysis of nanosized cell-derived membrane vesicles
AU - Nolte - t Hoen, Esther N M
AU - van der Vlist, Els J.
AU - Aalberts, Marian
AU - Mertens, Hendrik C H
AU - Bosch, Berend Jan
AU - Bartelink, Willem
AU - Mastrobattista, Enrico
AU - van Gaal, Ethlinn V B
AU - Stoorvogel, Willem
AU - Arkesteijn, Ger J.A. Arkesteijn
AU - Wauben, Marca H M
PY - 2012/7
Y1 - 2012/7
N2 - 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. From the Clinical Editor: The authors developed a fluorescence-based, high-resolution flow cytometric method for quantitative and qualitative analysis of nanosized cell-derived membrane vesicles that are increasingly recognized both as therapeutic vehicles and high-potential biomarkers for several diseases. A high throughput, easily available, and sensitive detection method such as the one discussed here is a critically important prerequisite for further refinements of this technology.
AB - 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. From the Clinical Editor: The authors developed a fluorescence-based, high-resolution flow cytometric method for quantitative and qualitative analysis of nanosized cell-derived membrane vesicles that are increasingly recognized both as therapeutic vehicles and high-potential biomarkers for several diseases. A high throughput, easily available, and sensitive detection method such as the one discussed here is a critically important prerequisite for further refinements of this technology.
KW - Biomarkers
KW - Cellular membrane vesicles
KW - Dendritic cells
KW - Exosomes
KW - Flow cytometry
KW - Microvesicles
KW - Nanovesicles
UR - https://www.scopus.com/pages/publications/84862677632
U2 - 10.1016/j.nano.2011.09.006
DO - 10.1016/j.nano.2011.09.006
M3 - Article
C2 - 22024193
AN - SCOPUS:84862677632
SN - 1549-9634
VL - 8
SP - 712
EP - 720
JO - Nanomedicine : nanotechnology, biology and medicine
JF - Nanomedicine : nanotechnology, biology and medicine
IS - 5
ER -