Validation of a scalable method using chromatography to isolate engineered extracellular vesicles carring igf1 derived from human umbilical cord perivascular cells for the treatment of experimental liver fibrosis

CANTERO, MARÍA JOSE; ATORRASAGASTI, CATALINA; FIORE, ESTEBAN; BUELONI, BÁRBARA; GARCIA, MARIANA; MAZZOLINI, GUILLERMO; DOMINGUEZ LUCIANA; BAYO, JUAN; YANNARELLI, GUSTAVO

Buenos Aires

Congreso; LXVI Reunión Científica Anual de la Sociedad Argentina de Investigación Clínica (SAIC); 2021

Sociedad Argentina de Investigación Clínica (SAIC)

Introduction: Extracellular vesicles (EVs) derived from human umbilical cord perivascular cells (HUCPVC) over-expressing IGF1 mediates therapeutic effect on liver fibrosis in mice. We aimed to validate these results using EVs isolated by affinity chromatography, a scalable method. Methods: HUCPVC were infected with adenoviruses codifying for human IGF1 (AdIGF1) or green fluorescence protein (AdGFP). Viability was determined by MTT assay. IGF1, TNF-a, and CD63 levels were determined by ELISA. EVs were isolated from HUCPVC supernatants by anion exchange chromatography and characterized by electron microscopy. Expression of pro-fibrogenic genes (Col1A2 and aSMA) on hepatic stellate cells (LX2) were determined by qPCR. Antifibrotic effect of EVs was determined in BALB/c mice with liver fibrosis (thioacetamide for 8 weeks). The treatments were administered on week 6 (Groups: Saline, EVs-AdIGFI or EVs-AdGFP, 3 doses, 15 mg/dose/mice every 5 days). Results: First, we found that HUCPVC infected with 2.5 to 30 MOI showed over-expression of IGF1, keep cell viability and exert an anti-inflammatory capacity on J774 macrophages revealed by decreased TNF-a expression (p