Identifier to cite or link to this item: http://hdl.handle.net/20.500.13003/11875
Purity Determines the Effect of Extracellular Vesicles Derived from Mesenchymal Stromal Cells
Identifiers
DOI: 10.3390/cells9020422
eISSN: 2073-4409
WOS ID: 000521944900163
Scopus EID: 2-s2.0-85093899243
PMID: 32059497
Embase PUI: L2003765431
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2020-02Document type
research articleCitation
Forteza-Genestra MA, Antich-Rossello M, Calvo J, Gaya A, Monjo M, Ramis JM. Purity Determines the Effect of Extracellular Vesicles Derived from Mesenchymal Stromal Cells. Cells. 2020 Feb;9(2):422.Abstract
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 Prot(w/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.
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https://dx.doi.org/10.3390/cells9020422Keywords
extracellular vesiclesultracentrifugation
size exclusion chromatography
ATDC-5 cell line
gene expression
collagen
human umbilical cord mesenchymal stromal cells
FBS
conditioned media
purity
MeSH
Umbilical CordExtracellular Vesicles
Metalloproteases
Gene Expression Regulation
Humans
Cells, Cultured
Decorin
Culture Media
Particle Size
Mesenchymal Stem Cells
Animals
Aggrecans
RNA, Messenger
Mice
DeCS
AnimalesMetaloproteasas
Medios de Cultivo
Agrecanos
Regulación de la Expresión Génica
Tamaño de la Partícula
Humanos
Células Cultivadas
Vesículas Extracelulares
Decorina
Células Madre Mesenquimatosas
ARN Mensajero
Ratones
Cordón Umbilical
This item appears in following Docusalut collections
Fundación Banco de Sangre y Tejidos de las Islas Baleares - FBSTIB > Comunicación científicaInstituto de Investigación Sanitaria Islas Baleares - IDISBA > Comunicación científica