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Mesenchymal Stem Cell Exosomes transfer microRNA and protect injured tubular epithelial cells.
Author(s) -
Borges Fernanda,
Schor Nestor,
Novaes Antonio
Publication year - 2015
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.29.1_supplement.670.8
Subject(s) - microvesicles , mesenchymal stem cell , microbiology and biotechnology , paracrine signalling , stem cell , microrna , exosome , extracellular vesicle , transfection , chemistry , biology , cell culture , receptor , biochemistry , gene , genetics
Bone Marrow Mesenchymal Stem Cells (BM‐MSCs) and Umbilical Cord Stem Cell (UCSC) protect the kidney against nephrotoxic insults through paracrine effects, possibly mediated by extracellular vesicles. Exosomes (EXOs) are up to 100nm extracellular vesicles that transfer mRNA and microRNA to the recipient cells, mediating the communication between them. MicroRNA 23a, 126 and 296 were shown to mediate the protective effect of stem cell microvesicles. The goal of the present study was to analyze the effects of EXOs from MSCs and UCSC on rat proximal tubular epithelial cells (RPTC) exposed to toxic insults. MSCs were maintained in culture conditions and every 3 days the culture medium was collected. EXOs were extracted through filtration in 0.1µm filter, followed by ultracentrifugation's (100,000g during 2h). 23a, 126 and 296 microRNAs from MSCs and EXOs were extracted and analyzed by real time‐PCR. RPTC cells were exposed to gentamicin (Genta, 2mM) in the presence or absence of exosomes (50 µg/ml) and proliferation was analyzed from 24 up to 96h through MTT assay. USCS and MSCs were instable transfected with EXOs marker GFP‐CD63, cultured on the top of an insert. On the bottom of the insert, tubular cells were treated with Genta. After 24h, the fluorescence of GFP was observed on tubular cells, suggesting that EXOs were taken up by tubular cells. Related to the endogenous control, the microRNA 126 was the main miRNA expressed in the UCSC‐EXO, while all of them were expressed equaly in MSC‐EXO. The treatment with UCSC‐EXO stimulated the proliferation of tubular cells exposed to Genta. Our results suggest that EXOs carrying microRNA from stem cells are taken up and protect tubular epithelial cells, mediating the proximal tubule regeneration. Supported by FAPESP, CNPq, CAPES and FOR