title: In vitro differentiation of porcine aortic vascular precursor cells to endothelial and vascular smooth muscle cells.
abstract: Recent findings suggest that progenitor and multipotent mesenchymal stromal cells (MSCs) are associated with vascular niches. Cells displaying mesenchymal properties and differentiating to whole components of a functional blood vessel, including endothelial and smooth muscle cells, can be defined as vascular stem cells (VSCs). Recently, we isolated a population of porcine aortic vascular precursor cells (pAVPCs), which have MSC- and pericyte-like properties. The aim of the present work was to investigate whether pAVPCs possess VSC-like properties and assess their differentiation potential toward endothelial and smooth muscle lineages. pAVPCs, maintained in a specific pericyte growth medium, were cultured in high-glucose DMEM + 10% FBS (long-term medium, LTM) or in human endothelial serum-free medium + 5% FBS and 50 ng/ml of hVEGF (endothelial differentiation medium, EDM). After 21 days of culture in LTM, pAVPCs showed an elongated fibroblast-like morphology, and they seem to organize in cord-like structures. qPCR analysis of smooth muscle markers [α-smooth muscle actin (α-SMA), calponin, and smooth muscle myosin (SMM) heavy chain] showed a significant increment of the transcripts, and immunofluorescence analysis confirmed the presence of α-SMA and SMM proteins. After 21 days of culture in EDM, pAVPCs displayed an endothelial cell-like morphology and revealed the upregulation of the expression of endothelial markers (CD31, vascular endothelial-cadherin, von Willebrand factor, and endothelial nitric oxide synthase) showing the CD31-typical pattern. In conclusion, pAVPCs could be defined as a VSC-like population considering that, if they are maintained in a specific pericyte medium, they express MSC markers, and they have, in addition to the classical mesenchymal trilineage differentiation potential, the capacity to differentiate in vitro toward the smooth muscle and the endothelial cell phenotypes.
authors.affiliation: Department of Veterinary Medical Sciences (DIMEVET) University of Bologna, Bologna, Italy;
authors.forename: Andrea
authors.initials: A
authors.lastname: Zaniboni
authors.affiliation: Department of Veterinary Medical Sciences (DIMEVET) University of Bologna, Bologna, Italy; chiara.bernardini5@unibo.it.
authors.forename: Chiara
authors.initials: C
authors.lastname: Bernardini
authors.affiliation: Department of Veterinary Medical Sciences (DIMEVET) University of Bologna, Bologna, Italy;
authors.forename: Martina
authors.initials: M
authors.lastname: Bertocchi
authors.affiliation: Department of Veterinary Medical Sciences (DIMEVET) University of Bologna, Bologna, Italy;
authors.forename: Augusta
authors.initials: A
authors.lastname: Zannoni
authors.affiliation: Stem Wave Institute for Tissue Healing (SWITH), Gruppo Villa Maria (GVM) Care and Research, Ettore Sansavini Health Science Foundation, Lugo (Ravenna), Italy; National Institute of Biostructures and Biosystems at the Department of Experimental, Diagnostic and Specialty Medicine, S. Orsola-Malpighi Hospital, University of Bologna, Bologna, Italy;
authors.forename: Francesca
authors.initials: F
authors.lastname: Bianchi
authors.affiliation: Department of Veterinary Medical Sciences (DIMEVET) University of Bologna, Bologna, Italy;
authors.forename: Giancarlo
authors.initials: G
authors.lastname: Avallone
authors.affiliation: Department of Veterinary Medical Sciences (DIMEVET) University of Bologna, Bologna, Italy;
authors.forename: Chiara
authors.initials: C
authors.lastname: Mangano
authors.affiliation: Department of Veterinary Medical Sciences (DIMEVET) University of Bologna, Bologna, Italy;
authors.forename: Giuseppe
authors.initials: G
authors.lastname: Sarli
authors.affiliation: Department of Pharmacy and Biotechnology (FaBiT), University of Bologna, Bologna, Italy; Health Sciences and Technologies, Interdepartmental Center for Industrial Research (HST-ICIR), University of Bologna, Bologna, Italy.
authors.forename: Laura
authors.initials: L
authors.lastname: Calzà
authors.affiliation: Department of Veterinary Medical Sciences (DIMEVET) University of Bologna, Bologna, Italy;
authors.forename: Maria Laura
authors.initials: ML
authors.lastname: Bacci
authors.affiliation: Department of Veterinary Medical Sciences (DIMEVET) University of Bologna, Bologna, Italy;
authors.forename: Monica
authors.initials: M
authors.lastname: Forni
mesh_terms: D000818:Animals; D001011:Aorta / Q000166:cytology / Q000502:physiology*; D002454:Cell Differentiation / Q000502:physiology*; D002478:Cells, Cultured; D004730:Endothelium, Vascular / Q000166:cytology / Q000502:physiology*; D006801:Humans; D009131:Muscle, Smooth, Vascular / Q000166:cytology / Q000502:physiology*; D032389:Myocytes, Smooth Muscle / Q000502:physiology*; D013234:Stem Cells / Q000502:physiology*; D013552:Swine
keywords: endothelial differentiation; mesenchymal stromal cells; pig animal model; smooth muscle differentiation; vascular stem cells