title: Human amniotic fluid stem cell differentiation along smooth muscle lineage.
abstract: Functional smooth muscle engineering requires isolation and expansion of smooth muscle cells (SMCs), and this process is particularly challenging for visceral smooth muscle tissue where progenitor cells have not been clearly identified. Herein we showed for the first time that efficient SMCs can be obtained from human amniotic fluid stem cells (hAFSCs). Clonal lines were generated from c-kit(+) hAFSCs. Differentiation toward SM lineage (SMhAFSCs) was obtained using a medium conditioned by PDGF-BB and TGF-β1. Molecular assays revealed higher level of α smooth muscle actin (α-SMA), desmin, calponin, and smoothelin in SMhAFSCs when compared to hAFSCs. Ultrastructural analysis demonstrated that SMhAFSCs also presented in the cytoplasm increased intermediate filaments, dense bodies, and glycogen deposits like SMCs. SMhAFSC metabolism evaluated via mass spectrometry showed higher glucose oxidation and an enhanced response to mitogenic stimuli in comparison to hAFSCs. Patch clamp of transduced hAFSCs with lentiviral vectors encoding ZsGreen under the control of the α-SMA promoter was performed demonstrating that SMhAFSCs retained a smooth muscle cell-like electrophysiological fingerprint. Eventually SMhAFSCs contractility was evident both at single cell level and on a collagen gel. In conclusion, we showed here that hAFSCs under selective culture conditions are able to give rise to functional SMCs.
authors.affiliation: 2UCL Institute of Child Health and Great Ormond St. Hospital for Children, 30 Guilford St., London WC1N 1EH, UK. p.decoppi@ucl.ac.uk.
authors.forename: Marco
authors.initials: M
authors.lastname: Ghionzoli
authors.forename: Andrea
authors.initials: A
authors.lastname: Repele
authors.forename: Laura
authors.initials: L
authors.lastname: Sartiani
authors.forename: Giulia
authors.initials: G
authors.lastname: Costanzi
authors.forename: Astrid
authors.initials: A
authors.lastname: Parenti
authors.forename: Valentina
authors.initials: V
authors.lastname: Spinelli
authors.forename: Anna L
authors.initials: AL
authors.lastname: David
authors.forename: Massimo
authors.initials: M
authors.lastname: Garriboli
authors.forename: Giorgia
authors.initials: G
authors.lastname: Totonelli
authors.forename: Jun
authors.initials: J
authors.lastname: Tian
authors.forename: Stelios T
authors.initials: ST
authors.lastname: Andreadis
authors.forename: Elisabetta
authors.initials: E
authors.lastname: Cerbai
authors.forename: Alessandro
authors.initials: A
authors.lastname: Mugelli
authors.forename: Antonio
authors.initials: A
authors.lastname: Messineo
authors.forename: Agostino
authors.initials: A
authors.lastname: Pierro
authors.forename: Simon
authors.initials: S
authors.lastname: Eaton
authors.forename: Paolo
authors.initials: P
authors.lastname: De Coppi
mesh_terms: D000199:Actins / Q000235:genetics / Q000378:metabolism; D000200:Action Potentials; D000653:Amniotic Fluid / Q000166:cytology*; D002135:Calcium-Binding Proteins / Q000235:genetics / Q000378:metabolism; D002454:Cell Differentiation; D019070:Cell Lineage; D017077:Culture Media, Conditioned / Q000494:pharmacology; D003593:Cytoplasm / Q000378:metabolism / Q000648:ultrastructure; D003598:Cytoskeletal Proteins / Q000235:genetics / Q000378:metabolism; D003893:Desmin / Q000235:genetics / Q000378:metabolism; D053686:Fetal Stem Cells / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism / Q000502:physiology; D005947:Glucose / Q000378:metabolism; D006003:Glycogen / Q000378:metabolism; D006801:Humans; D008840:Microfilament Proteins / Q000235:genetics / Q000378:metabolism; D039902:Multipotent Stem Cells / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism / Q000502:physiology; D009124:Muscle Proteins / Q000235:genetics / Q000378:metabolism; D032389:Myocytes, Smooth Muscle / Q000166:cytology* / Q000378:metabolism / Q000502:physiology; D010982:Platelet-Derived Growth Factor / Q000494:pharmacology; D016212:Transforming Growth Factor beta / Q000494:pharmacology; D000096985:Calponins
chemical_list: D000199:Actins; D002135:Calcium-Binding Proteins; D017077:Culture Media, Conditioned; D003598:Cytoskeletal Proteins; D003893:Desmin; D005947:Glucose; D006003:Glycogen; D008840:Microfilament Proteins; D009124:Muscle Proteins; D010982:Platelet-Derived Growth Factor; D016212:Transforming Growth Factor beta; D000096985:Calponins; C100762:SMTN protein, human
keywords: fetal cells; multipotent; myogenic; regenerative medicine; tissue engineering