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oai:repisalud.isciii.es:20.500.12105/229432024-11-28T21:42:46Zcom_20.500.12105_15322com_20.500.12105_2051col_20.500.12105_16967

00925njm 22002777a 4500 dc Forteza-Genestra, Maria Antonia author Antich-Rosselló, Miquel author Calvo Benito, Javier author Gayà, Antoni author Monjo, Marta author Ramis, Joana Maria author 2020-02 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. 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. http://hdl.handle.net/20.500.13003/11875 https://hdl.handle.net/20.500.12105/22943 32059497 10.3390/cells9020422 2073-4409 Cells 2-s2.0-85093899243 521944900163 L2003765431 Extracellular vesicles Ultracentrifugation Size exclusion chromatography ATDC-5 cell line Gene expression Collagen Human umbilical cord mesenchymal stromal cells FBS Conditioned media Purity Purity Determines the Effect of Extracellular Vesicles Derived from Mesenchymal Stromal Cells