Comparison study of three mesenchymal stem cells-derived extracellular vesicles sources for experimental therapy in liver fibrosis.

DOMÍNGUEZ LM ; ALBORNOZ M; BAYO J; CANTERO MJ; BUELONI B; ATORRASAGASTI C; BIANI C; LUZZANI C; GARCIA M; YANNARELLI G; FIORE E; MAZZOLINI G

Congreso; Reunión Conjunta SAIC SAI&FAIC SAFIS 2022; 2022

Introduction: Liver cirrhosis involves chronic damage, wound healing and fibrogenic processes. Mesenchymal stem cells (MSC)-derived extracellular vesicles (EVs) are an interesting therapeutic option for regenerative medicine. Aim: To compare EVs derived from different clinical relevant sources of MSC as therapeutic tool for liver fibrosis. Methods: EVs were isolated by ion exchange chromatography from supernatants of adipose tissue MSCs (ASC-EV), induced pluripotent stem cells-derived MSCs- (iMSC-EV), and umbilical cord perivascular cells (HUCPVC-EV). EVs isolation was confirmed by protein (BCA) and CD63 (ELISA) quantification and markers expression (CD9, CD81) by flow cytometry. In vitro effects of EVs on hepatic stellate cells (HSC) pro-fibrogenic genes expression (Col1A2 and a-SMA) were evaluated in CFSC-G2 cell line by qPCR. Antifibrotic effect of EVs was determined in experimental mice model of liver fibrosis (thioacetamide for 8 weeks in BALB/c mice). On week 6, ASC-EV, iMSCs-EV and HUCPVC-EV were i.v. injected every 5 days for a total of 3 doses. At week 8, animals were sacrificed, and liver samples analyzed. Collagen deposition was measured by Sirius red staining; immunohistochemistry for PCNA proliferating cells, and a-SMA (a-smooth muscle actin) for activated HSC was performed. Results: The three MSC source produce significant amount of EVs with typical surface markers CD9, CD63 and CD81. In vitro HSC incubation with EVs from different MSC sources down-regulates in similar levels the Col1a2 (p