Protocol for cross-platform characterization of human and murine extracellular vesicles and particles

Linda Bojmar; Han Sang Kim; Kei Sugiura; Søren Heissel; Serena Lucotti; Michele Cioffi; Kofi Ennu Johnson; Leona Cohen-Gould; Haiying Zhang; Henrik Molina; Irina R. Matei; David Lyden; Ayuko Hoshino

STAR Protocols (Mar 2024)

Protocol for cross-platform characterization of human and murine extracellular vesicles and particles

Linda Bojmar,
Han Sang Kim,
Kei Sugiura,
Søren Heissel,
Serena Lucotti,
Michele Cioffi,
Kofi Ennu Johnson,
Leona Cohen-Gould,
Haiying Zhang,
Henrik Molina,
Irina R. Matei,
David Lyden,
Ayuko Hoshino

Affiliations

Linda Bojmar: Children’s Cancer and Blood Foundation Laboratories, Departments of Pediatrics, and Cell and Developmental Biology, Drukier Institute for Children’s Health, Meyer Cancer Center, Weill Cornell Medicine, New York, NY, USA; Department of Biomedical and Clinical Sciences, Linköping University, Linköping, Sweden
Han Sang Kim: Children’s Cancer and Blood Foundation Laboratories, Departments of Pediatrics, and Cell and Developmental Biology, Drukier Institute for Children’s Health, Meyer Cancer Center, Weill Cornell Medicine, New York, NY, USA; Yonsei Cancer Center, Division of Medical Oncology, Department of Internal Medicine, Graduate School of Medical Science, Brain Korea 21 FOUR Project, Yonsei University College of Medicine, Seoul, Korea
Kei Sugiura: Research Center for Advanced Science and Technology, The University of Tokyo, Tokyo, Japan; School of Life Science and Technology, Tokyo Institute of Technology, Yokohama, Japan; Graduate School of Pharmaceutical Sciences, The University of Tokyo, Tokyo, Japan
Søren Heissel: Proteomics Resource Center, The Rockefeller University, New York, NY, USA
Serena Lucotti: Children’s Cancer and Blood Foundation Laboratories, Departments of Pediatrics, and Cell and Developmental Biology, Drukier Institute for Children’s Health, Meyer Cancer Center, Weill Cornell Medicine, New York, NY, USA
Michele Cioffi: Children’s Cancer and Blood Foundation Laboratories, Departments of Pediatrics, and Cell and Developmental Biology, Drukier Institute for Children’s Health, Meyer Cancer Center, Weill Cornell Medicine, New York, NY, USA
Kofi Ennu Johnson: Children’s Cancer and Blood Foundation Laboratories, Departments of Pediatrics, and Cell and Developmental Biology, Drukier Institute for Children’s Health, Meyer Cancer Center, Weill Cornell Medicine, New York, NY, USA; Tri-Institutional PhD Program in Computational Biology and Medicine, New York, NY, USA
Leona Cohen-Gould: Microscopy & Image Analysis, Core Facilities, Weill Cornell Medicine, New York, NY, USA
Haiying Zhang: Children’s Cancer and Blood Foundation Laboratories, Departments of Pediatrics, and Cell and Developmental Biology, Drukier Institute for Children’s Health, Meyer Cancer Center, Weill Cornell Medicine, New York, NY, USA
Henrik Molina: Proteomics Resource Center, The Rockefeller University, New York, NY, USA
Irina R. Matei: Children’s Cancer and Blood Foundation Laboratories, Departments of Pediatrics, and Cell and Developmental Biology, Drukier Institute for Children’s Health, Meyer Cancer Center, Weill Cornell Medicine, New York, NY, USA; Corresponding author
David Lyden: Children’s Cancer and Blood Foundation Laboratories, Departments of Pediatrics, and Cell and Developmental Biology, Drukier Institute for Children’s Health, Meyer Cancer Center, Weill Cornell Medicine, New York, NY, USA; Corresponding author
Ayuko Hoshino: Children’s Cancer and Blood Foundation Laboratories, Departments of Pediatrics, and Cell and Developmental Biology, Drukier Institute for Children’s Health, Meyer Cancer Center, Weill Cornell Medicine, New York, NY, USA; Research Center for Advanced Science and Technology, The University of Tokyo, Tokyo, Japan; School of Life Science and Technology, Tokyo Institute of Technology, Yokohama, Japan; Corresponding author

Journal volume & issue: Vol. 5, no. 1
p. 102754

Abstract

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Summary: Characterization of isolated extracellular vesicles and particles (EVPs) is crucial for determining functions and biomarker potential. Here, we present a protocol to analyze size, number, morphology, and EVP protein cargo and to validate EVP proteins in both humans and mice. We describe steps for nanoparticle tracking analysis, transmission electron microscopy, single-EVP immunodetection, EVP proteomic mass spectrometry and bioinformatic analysis, and EVP protein validation by ExoELISA and western blot analysis. This allows for EVP cross-validation across different platforms.For complete details on the use and execution of this protocol, please refer to Hoshino et al.1 : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.

Published in STAR Protocols

ISSN: 2666-1667 (Online)
Publisher: Elsevier
Country of publisher: United States
LCC subjects: Science: Science (General)
Website: https://star-protocols.cell.com/

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