title: First Characterization of Human Amniotic Fluid Stem Cell Extracellular Vesicles as a Powerful Paracrine Tool Endowed with Regenerative Potential.
abstract: Human amniotic fluid stem cells (hAFS) have shown a distinct secretory profile and significant regenerative potential in several preclinical models of disease. Nevertheless, little is known about the detailed characterization of their secretome. Herein we show for the first time that hAFS actively release extracellular vesicles (EV) endowed with significant paracrine potential and regenerative effect. c-KIT+hAFS were isolated from leftover samples of amniotic fluid from prenatal screening and stimulated to enhance EV release (24 hours 20% O2versus 1% O2preconditioning). The capacity of the c-KIT+hAFS-derived EV (hAFS-EV) to induce proliferation, survival, immunomodulation, and angiogenesis were investigated in vitro and in vivo. The hAFS-EV regenerative potential was also assessed in a model of skeletal muscle atrophy (HSA-Cre, SmnF7/F7mice), in which mouse AFS transplantation was previously shown to enhance muscle strength and survival. hAFS secreted EV ranged from 50 up to 1,000 nm in size. In vitro analysis defined their role as biological mediators of regenerative, paracrine effects while their modulatory role in decreasing skeletal muscle inflammation in vivo was shown for the first time. Hypoxic preconditioning significantly induced the enrichment of exosomes endowed with regenerative microRNAs within the hAFS-EV. In conclusion, this is the first study showing that c-KIT+hAFS dynamically release EV endowed with remarkable paracrine potential, thus representing an appealing tool for future regenerative therapy. Stem Cells Translational Medicine 2017;6:1340-1355.
authors.affiliation: Regenerative Medicine Laboratory, Department of Experimental Medicine, University of Genova, Genova, Italy.
authors.forename: Carolina
authors.initials: C
authors.lastname: Balbi
authors.affiliation: Stem Cells and Regenerative Medicine Laboratory, Fondazione Istituto di Ricerca Pediatrica Città della Speranza, Padova, Italy.
authors.forename: Martina
authors.initials: M
authors.lastname: Piccoli
authors.affiliation: Laboratory of Molecular and Cellular Cardiology, CardioCentro Ticino Foundation_CCT, Lugano Switzerland.
authors.forename: Lucio
authors.initials: L
authors.lastname: Barile
authors.affiliation: Regenerative Medicine Laboratory, Department of Experimental Medicine, University of Genova, Genova, Italy.
authors.forename: Andrea
authors.initials: A
authors.lastname: Papait
authors.affiliation: Drug Discovery and Development Department, IIT-Fondazione Istituto Italiano di Tecnologia, Genova, Italy.
authors.forename: Andrea
authors.initials: A
authors.lastname: Armirotti
authors.affiliation: Regenerative Medicine Laboratory, Department of Experimental Medicine, University of Genova, Genova, Italy.
authors.forename: Elisa
authors.initials: E
authors.lastname: Principi
authors.affiliation: Molecular Pathology Unit, IRCCS AOU San Martino - IST National Institute for Cancer Research, Genova, Italy.
authors.forename: Daniele
authors.initials: D
authors.lastname: Reverberi
authors.affiliation: Veterinary Medicine Department, University of Perugia, Perugia, Italy.
authors.forename: Luisa
authors.initials: L
authors.lastname: Pascucci
authors.affiliation: Molecular Biology Laboratory, IRCCS Istituto Giannina Gaslini, Genova, Italy.
authors.forename: Pamela
authors.initials: P
authors.lastname: Becherini
authors.affiliation: Molecular Biology Laboratory, IRCCS Istituto Giannina Gaslini, Genova, Italy.
authors.forename: Luigi
authors.initials: L
authors.lastname: Varesio
authors.affiliation: Human Genetics Laboratory, E.O. Ospedali Galliera, Genova, Italy.
authors.forename: Massimo
authors.initials: M
authors.lastname: Mogni
authors.affiliation: Human Genetics Laboratory, E.O. Ospedali Galliera, Genova, Italy.
authors.forename: Domenico
authors.initials: D
authors.lastname: Coviello
authors.affiliation: Drug Discovery and Development Department, IIT-Fondazione Istituto Italiano di Tecnologia, Genova, Italy.
authors.forename: Tiziano
authors.initials: T
authors.lastname: Bandiera
authors.affiliation: Stem Cells and Regenerative Medicine Laboratory, Fondazione Istituto di Ricerca Pediatrica Città della Speranza, Padova, Italy.|Department of Woman and Child Health, University of Padova, Padova, Italy.
authors.forename: Michela
authors.initials: M
authors.lastname: Pozzobon
authors.affiliation: Regenerative Medicine Laboratory, Department of Experimental Medicine, University of Genova, Genova, Italy.
authors.forename: Ranieri
authors.initials: R
authors.lastname: Cancedda
authors.affiliation: Regenerative Medicine Laboratory, Department of Experimental Medicine, University of Genova, Genova, Italy.
authors.forename: Sveva
authors.initials: S
authors.lastname: Bollini
mesh_terms: D000653:Amniotic Fluid / Q000166:cytology*; D000818:Animals; D002454:Cell Differentiation; D049109:Cell Proliferation; D000067128:Extracellular Vesicles / Q000378:metabolism*; D006801:Humans; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D035683:MicroRNAs; D018482:Muscle, Skeletal / Q000166:cytology; D009133:Muscular Atrophy / Q000628:therapy; D033581:Stem Cell Transplantation; D013234:Stem Cells / Q000166:cytology / Q000502:physiology
chemical_list: D035683:MicroRNAs
keywords: Apoptosis; Exosomes; Extracellular vesicles; Fetal stem cells; Paracrine communication; Proliferation; Tissue regeneration; miRNA