Proteomic analysis of mesenchymal stromal cell-derived extracellular vesicles and reconstructed membrane particles

Hector Tejeda-Mora, Leticia G. Leon, Jeroen Demmers, Carla C. Baan, Marlies E.J. Reinders, Bertram Bleck, Eleuterio Lombardo, Ana Merino, Martin J. Hoogduijn*

*Corresponding author for this work

Research output: Contribution to journalArticleAcademicpeer-review

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Extracellular vesicles (EV) derived from mesenchymal stromal cells (MSC) are a potential therapy for immunological and degenerative diseases. However, large-scale production of EV free from contamination by soluble proteins is a major challenge. The generation of particles from isolated membranes of MSC, membrane particles (MP), may be an alternative to EV. In the present study we generated MP from the membranes of lysed MSC after removal of the nuclei. The yield of MP per MSC was 1 × 105 times higher than EV derived from the same number of MSC. To compare the proteome of MP and EV, proteomic analysis of MP and EV was performed. MP contained over 20 times more proteins than EV. The proteins present in MP evidenced a multi-organelle origin of MP. The projected function of the proteins in EV and MP was very different. Whilst proteins in EV mainly play a role in extracellular matrix organization, proteins in MP were interconnected in diverse molecular pathways, including protein synthesis and degradation pathways and demon-strated enzymatic activity. Treatment of MSC with IFNγ led to a profound effect on the protein make up of EV and MP, demonstrating the possibility to modify the phenotype of EV and MP through modification of parent MSC. These results demonstrate that MP are an attractive alternative to EV for the development of potential therapies. Functional studies will have to demonstrate therapeutic efficacy of MP in preclinical disease models.

Original languageEnglish
Article number12935
JournalInternational Journal of Molecular Sciences
Issue number23
Publication statusPublished - 29 Nov 2021

Bibliographical note

Funding Information:
This collaboration project is co-funded by the PPP Allowance made available by Health~Holland, Top Sector Life Sciences & Health, to stimulate public-private partnerships.

Publisher Copyright:
© 2021 by the authors.


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