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Isolation and characterization of microvesicles from mesenchymal stem cells.

Journal article
Authors M Rezaa Mohammadi
Milad Riazifar
Egest J Pone
Ashish Yeri
Kendall Van Keuren-Jensen
Cecilia Lässer
Jan Lötvall
Weian Zhao
Published in Methods (San Diego, Calif.)
ISSN 1095-9130
Publication year 2019
Published at Krefting Research Centre
Language en
Links dx.doi.org/10.1016/j.ymeth.2019.10....
www.ncbi.nlm.nih.gov/entrez/query.f...
Subject categories Other Medical Sciences not elsewhere specified

Abstract

Mesenchymal stem or stromal cells are currently under clinical investigation for multiple diseases. While their mechanism of action is still not fully elucidated, vesicles secreted by MSCs are believed to recapitulate their therapeutic potentials to some extent. Microvesicles (MVs), also called as microparticles or ectosome, are among secreted vesicles that could transfer cytoplasmic cargo, including RNA and proteins, from emitting (source) cells to recipient cells. Given the importance of MVs, we here attempted to establish a method to isolate and characterize MVs secreted from unmodified human bone marrow derived MSCs (referred to as native MSCs, and their microvesicles as Native-MVs) and IFNγ stimulated MSCs (referred to as IFNγ-MSCs, and their microvesicles as IFNγ-MVs). We first describe an ultracentrifugation technique to isolate MVs from the conditioned cell culture media of MSCs. Next, we describe characterization and quality control steps to analyze the protein and RNA content of MVs. Finally, we examined the potential of MVs to exert immunomodulatory effects through induction of regulatory T cells (Tregs). Secretory vesicles from MSCs are promising alternatives for cell therapy with applications in drug delivery, regenerative medicine, and immunotherapy.

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