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

Hoen, Esther N.M. Nolte-'t; van der Vlist, Els J.; Aalberts, Marian; Mertens, Hendrik C.H.; Bosch, Berend Jan; Bartelink, Willem; Mastrobattista, Enrico; van Gaal, Ethlinn V.B.; Stoorvogel, Willem; Arkesteijn, Ger J.A.; Wauben, Marca H.M.

doi:10.1016/j.nano.2011.09.006

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

Esther N.M. Nolte-'t Hoen, PhD
- Department of Biochemistry & Cell Biology, Faculty of Veterinary Medicine, Utrecht University, Utrecht, The Netherlands
Els J. van der Vlist, MSc
- Department of Biochemistry & Cell Biology, Faculty of Veterinary Medicine, Utrecht University, Utrecht, The Netherlands
Marian Aalberts, MSc
- Department of Biochemistry & Cell Biology, Faculty of Veterinary Medicine, Utrecht University, Utrecht, The Netherlands
Hendrik C.H. Mertens
- Department of Biochemistry & Cell Biology, Faculty of Veterinary Medicine, Utrecht University, Utrecht, The Netherlands
Berend Jan Bosch, PhD
- Department of Virology, Faculty of Veterinary Medicine, Utrecht University, Utrecht, The Netherlands
Willem Bartelink
- Department of Virology, Faculty of Veterinary Medicine, Utrecht University, Utrecht, The Netherlands
Enrico Mastrobattista, PhD
- Department of Pharmaceutics, Utrecht Institute for Pharmaceutical Sciences (UIPS), Utrecht University, Utrecht, The Netherlands
Ethlinn V.B. van Gaal, PhD
- Department of Pharmaceutics, Utrecht Institute for Pharmaceutical Sciences (UIPS), Utrecht University, Utrecht, The Netherlands
Willem Stoorvogel, PhD
- Department of Biochemistry & Cell Biology, Faculty of Veterinary Medicine, Utrecht University, Utrecht, The Netherlands
Ger J.A. Arkesteijn, PhD
- Department of Immunology, Faculty of Veterinary Medicine, Utrecht University, Utrecht, The Netherlands
Marca H.M. Wauben, PhD
- Department of Biochemistry & Cell Biology, Faculty of Veterinary Medicine, Utrecht University, Utrecht, The Netherlands
- Corresponding author: Faculty of Veterinary Medicine, Dept. of Biochemistry & Cell Biology, Utrecht University, Yalelaan 2, 3584CM Utrecht, The Netherlands.

Received 5 July 2011; accepted 23 September 2011. published online 24 October 2011.
Corrected Proof

Abstract

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.