Next Article in Journal
Asbestos and Intrahepatic Cholangiocarcinoma
Next Article in Special Issue
Extracellular Vesicles Derived from Induced Pluripotent Stem Cells Promote Renoprotection in Acute Kidney Injury Model
Previous Article in Journal
Intravenous Administration of Allogenic Cell-Derived Microvesicles of Healthy Origins Defend Against Atherosclerotic Cardiovascular Disease Development by a Direct Action on Endothelial Progenitor Cells
Previous Article in Special Issue
Potential Applications of Extracellular Vesicles in Solid Organ Transplantation
Open AccessArticle

Purity Determines the Effect of Extracellular Vesicles Derived from Mesenchymal Stromal Cells

1
Cell Therapy and Tissue Engineering Group, Research Institute on Health Sciences (IUNICS), University of the Balearic Islands, Ctra Valldemossa km 7.5, 07122 Palma, Spain
2
Health Research Institute of the Balearic Islands (IdISBa), 07120 Palma, Spain
3
Fundació Banc de Sang i Teixits de les Illes Balears (FBSTIB), 07004 Palma, Spain
*
Authors to whom correspondence should be addressed.
Both authors contributed equally to this work.
Cells 2020, 9(2), 422; https://doi.org/10.3390/cells9020422 (registering DOI)
Received: 15 January 2020 / Revised: 5 February 2020 / Accepted: 10 February 2020 / Published: 12 February 2020
(This article belongs to the Special Issue Therapeutic Applications of Extracellular Vesicles)
Extracellular vesicles (EVs) have been recently identified as vital components of cell-based therapies based on the observation that conditioned media from cultured stromal cells reproduce some of the beneficial effects of intact cells. In order to obtain clinically active EVs derived from Mesenchymal Stromal Cells (MSCs) different procedures have been reported in the literature. Usually, non-confluent cells are incubated with culture medium for 48 h either with EV-depleted Fetal Bovine Serum (FBS) or without FBS. Our aim was to compare the effects of EVs isolated by ultracentrifugation from human umbilical cord MSC conditioned media obtained using these two conditions: with EV-depleted FBS (UC) or without FBS (UCw/o) on the mRNA expression levels of extracellular matrix related genes using the mouse chondrogenic cell line ATDC-5. We observed a deleterious effect on chondrogenic cells treated with UCw/o, showing higher mRNA expression levels of different metalloproteinases and decorin (Dcn) and lower collagen (Col1a1 and Col2a1) and aggrecan (Acan) mRNA levels. To elucidate whether this deleterious effect was induced by the EVs or by any proteins co-purified in the EV pellet, we used size exclusion chromatography (SEC) to further purify the EV pellet, obtaining an EV enriched fraction (EV or EVw/o) and a protein enriched fraction (Prot or Protw/o). Our results pointed that the negative effect on the chondrogenic cell line was due to the contaminant proteins coisolated with the EVs by ultracentrifugation and not from the EVs themselves. Thus, these results highlight the importance of working with well purified EV preparations to specifically achieve their therapeutic effect.
Keywords: extracellular vesicles; ultracentrifugation; size exclusion chromatography; ATDC-5 cell line; gene expression; collagen; human umbilical cord mesenchymal stromal cells; FBS; conditioned media; purity extracellular vesicles; ultracentrifugation; size exclusion chromatography; ATDC-5 cell line; gene expression; collagen; human umbilical cord mesenchymal stromal cells; FBS; conditioned media; purity
MDPI and ACS Style

Forteza-Genestra, M.A.; Antich-Rosselló, M.; Calvo, J.; Gayà, A.; Monjo, M.; Ramis, J.M. Purity Determines the Effect of Extracellular Vesicles Derived from Mesenchymal Stromal Cells. Cells 2020, 9, 422.

Show more citation formats Show less citations formats
Note that from the first issue of 2016, MDPI journals use article numbers instead of page numbers. See further details here.

Article Access Map by Country/Region

1
Back to TopTop