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Isolation and characterization of extracellular vesicles from Caenorhabditis elegans for multi-omic analysis.

By Joshua C Russell, Gennifer E Merrihew, Julia E Robbins, Nadia Postupna, Tyek-Kyun Kim, Alexandra Golubeva, Ayush Noori, Kai Wang, C Dirk Keene, Michael J. MacCoss, Matt Kaeberlein

Posted 23 Nov 2018
bioRxiv DOI: 10.1101/476226

Cells from bacteria to human release vesicles into their extracellular environment. These extracellular vesicles (EVs) contain multiple classes of molecules, including nucleic acids, proteins, and lipids. The isolation and analysis of EV cargos from mammalian cell culture and liquid biopsy samples has become a powerful approach for uncovering the messages that are packaged into these organelles. However, this type of research is not yet possible for invertebrate model systems, because methods have yet to be developed to obtain sufficient amounts of pure EVs. Here we report a robust and reproducible procedure to isolate EVs from Caenorhabditis elegans with yields similar to those obtained from human cell culture. Through nanoparticle tracking, transmission electron microscopy, flow cytometry, mass spectrometry, RNAseq, and immunoaffinity analysis we provide the first ever detailed characterization of C. elegans EV composition and demonstrate that C. elegans EVs share fundamentally similar properties with their mammalian counterparts. These include vesicle size, enrichment for lipid rafts, and similar types of RNA and protein cargos. This ability of isolate pure EVs on a scale amenable to multiple types of downstream analyses permits, for the first time, multi-omics characterization of EV cargos in an invertebrate model system.

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