Purity Determines the Effect of Extracellular Vesicles Derived from Mesenchymal Stromal Cells
| Autor: | Javier Calvo, Marta Monjo, Miquel Antich-Rosselló, Maria Antònia Forteza-Genestra, Joana M. Ramis, Antoni Gayà |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 2020 |
| Předmět: |
collagen
medios de cultivo Decorin Cell humanos human umbilical cord mesenchymal stromal cells metaloproteasas regulación de la expresión génica FBS Umbilical Cord Extracellular matrix Mice ultracentrifugation purity Aggrecans lcsh:QH301-705.5 decorina Cells Cultured Chemistry size exclusion chromatography General Medicine Cell biology ARN medicine.anatomical_structure ATDC-5 cell line conditioned media extracellular vesicles Stromal cell Cells Article cordón umbilical medicine Animals Humans RNA Messenger Particle Size tamaño de partículas Aggrecan Mesenchymal stem cell Mesenchymal Stem Cells agrecanos Culture Media lcsh:Biology (General) Gene Expression Regulation Cell culture Metalloproteases gene expression RNA animales células ratones Fetal bovine serum |
| Zdroj: | Cells Cells, Vol 9, Iss 2, p 422 (2020) Volume 9 Issue 2 |
| Popis: | 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. This research was funded by Instituto de Salud Carlos III, co-funded by the ESF European Social Fund and the ERDF European Regional Development Fund (grant number MS16/00124; CP16/00124), Ministerio de Economia y Competividad (IEDI-2017-00941), and the Direccio General d'Investigacio, Conselleria d'Investigacio, Govern Balear (grant number FPI/2046/2017). |
| Databáze: | OpenAIRE |
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