Proteomic profiling of mesenchymal stem cell-derived extracellular vesicles: Impact of isolation methods on protein cargo.

Autor: Abyadeh M; Department of Stem Cells and Developmental Biology, Cell Science Research Center Royan Institute for Stem Cell Biology and Technology, ACECR Tehran Iran., Mirshahvaladi S; Macquarie Medical School, School of Medicine Health and Human Sciences, Macquarie University Sydney New South Wales Australia., Kashani SA; Department of Stem Cells and Developmental Biology, Cell Science Research Center Royan Institute for Stem Cell Biology and Technology, ACECR Tehran Iran.; Motor Neuron Disease Research Centre, Faculty of Medicine, Health and Human Sciences Macquarie University Sydney New South Wales Australia., Paulo JA; Department of Cell Biology Harvard Medical School Boston Massachusetts USA., Amirkhani A; Australian Proteome Analysis Facility Macquarie University Sydney New South Wales Australia., Mehryab F; Advanced Therapy Medicinal Product Technology Development Center, Cell Science Research Center Royan Institute for Stem Cell Biology and Technology, ACECR Tehran Iran., Seydi H; Advanced Therapy Medicinal Product Technology Development Center, Cell Science Research Center Royan Institute for Stem Cell Biology and Technology, ACECR Tehran Iran.; Department of Biology University of Science and Culture Tehran Iran., Moradpour N; Department of Biology University of Science and Culture Tehran Iran., Jodeiryjabarzade S; Department of Biology University of Science and Culture Tehran Iran., Mirzaei M; Macquarie Medical School, School of Medicine Health and Human Sciences, Macquarie University Sydney New South Wales Australia., Gupta V; Macquarie Medical School, School of Medicine Health and Human Sciences, Macquarie University Sydney New South Wales Australia., Shekari F; Department of Stem Cells and Developmental Biology, Cell Science Research Center Royan Institute for Stem Cell Biology and Technology, ACECR Tehran Iran.; Advanced Therapy Medicinal Product Technology Development Center, Cell Science Research Center Royan Institute for Stem Cell Biology and Technology, ACECR Tehran Iran., Salekdeh GH; Faculty of Natural Sciences Macquarie University North Ryde New South Wales Australia.
Jazyk: angličtina
Zdroj: Journal of extracellular biology [J Extracell Biol] 2024 Jun 06; Vol. 3 (6), pp. e159. Date of Electronic Publication: 2024 Jun 06 (Print Publication: 2024).
DOI: 10.1002/jex2.159
Abstrakt: Extracellular vesicles (EVs) are nanosized vesicles with a lipid bilayer that are secreted by cells and play a critical role in cell-to-cell communication. Despite the promising reports regarding their diagnostic and therapeutic potential, the utilization of EVs in the clinical setting is limited due to insufficient information about their cargo and a lack of standardization in isolation and analysis methods. Considering protein cargos in EVs as key contributors to their therapeutic potency, we conducted a tandem mass tag (TMT) quantitative proteomics analysis of three subpopulations of mesenchymal stem cell (MSC)-derived EVs obtained through three different isolation techniques: ultracentrifugation (UC), high-speed centrifugation (HS), and ultracentrifugation on sucrose cushion (SU). Subsequently, we checked EV marker expression, size distribution, and morphological characterization, followed by bioinformatic analysis. The bioinformatic analysis of the proteome results revealed that these subpopulations exhibit distinct molecular and functional characteristics. The choice of isolation method impacts the proteome of isolated EVs by isolating different subpopulations of EVs. Specifically, EVs isolated through the high-speed centrifugation (HS) method exhibited a higher abundance of ribosomal and mitochondrial proteins. Functional apoptosis assays comparing isolated mitochondria with EVs isolated through different methods revealed that HS-EVs, but not other EVs, induced early apoptosis in cancer cells. On the other hand, EVs isolated using the sucrose cushion (SU) and ultracentrifugation (UC) methods demonstrated a higher abundance of proteins primarily involved in the immune response, cell-cell interactions and extracellular matrix interactions. Our analyses unveil notable disparities in proteins and associated biological functions among EV subpopulations, underscoring the importance of meticulously selecting isolation methods and resultant EV subpopulations based on the intended application.
Competing Interests: The authors have no conflicts of interest to disclose.
(© 2024 The Author(s). Journal of Extracellular Biology published by Wiley Periodicals LLC on behalf of International Society for Extracellular Vesicles.)
Databáze: MEDLINE
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