title: Expansion and characterization of neonatal cardiac pericytes provides a novel cellular option for tissue engineering in congenital heart disease.
abstract: BACKGROUND
  Living grafts produced by combining autologous heart-resident stem/progenitor cells and tissue engineering could provide a new therapeutic option for definitive correction of congenital heart disease. The aim of the study was to investigate the antigenic profile, expansion/differentiation capacity, paracrine activity, and pro-angiogenic potential of cardiac pericytes and to assess their engrafting capacity in clinically certified prosthetic grafts.
  
  
  RESULTS
  CD34(pos) cells, negative for the endothelial markers CD31 and CD146, were identified by immunohistochemistry in cardiac leftovers from infants and children undergoing palliative repair of congenital cardiac defects. Following isolation by immunomagnetic bead-sorting and culture on plastic in EGM-2 medium supplemented with growth factors and serum, CD34(pos)/CD31(neg) cells gave rise to a clonogenic, highly proliferative (>20 million at P5), spindle-shape cell population. The following populations were shown to expresses pericyte/mesenchymal and stemness markers. After exposure to differentiation media, the expanded cardiac pericytes acquired markers of vascular smooth muscle cells, but failed to differentiate into endothelial cells or cardiomyocytes. However, in Matrigel, cardiac pericytes form networks and enhance the network capacity of endothelial cells. Moreover, they produce collagen-1 and release chemo-attractants that stimulate the migration of c-Kit(pos) cardiac stem cells. Cardiac pericytes were then seeded onto clinically approved xenograft scaffolds and cultured in a bioreactor. After 3 weeks, fluorescent microscopy showed that cardiac pericytes had penetrated into and colonized the graft.
  
  
  CONCLUSIONS
  These findings open new avenues for cellular functionalization of prosthetic grafts to be applied in reconstructive surgery of congenital heart disease.
authors.affiliation: Division of Experimental Cardiovascular Medicine, Bristol Heart Institute, University of Bristol, United Kingdom (E.A., I.R.A., H.L.S., F.R., G.M., S.C.S., J.R., V.V.A., O.O.I., S.S., A.O., P.M.).
authors.forename: Elisa
authors.initials: E
authors.lastname: Avolio
authors.affiliation: Division of Experimental Cardiovascular Medicine, Bristol Heart Institute, University of Bristol, United Kingdom (E.A., I.R.A., H.L.S., F.R., G.M., S.C.S., J.R., V.V.A., O.O.I., S.S., A.O., P.M.).
authors.forename: Iker
authors.initials: I
authors.lastname: Rodriguez-Arabaolaza
authors.affiliation: Division of Experimental Cardiovascular Medicine, Bristol Heart Institute, University of Bristol, United Kingdom (E.A., I.R.A., H.L.S., F.R., G.M., S.C.S., J.R., V.V.A., O.O.I., S.S., A.O., P.M.).
authors.forename: Helen L
authors.initials: HL
authors.lastname: Spencer
authors.affiliation: Division of Experimental Cardiovascular Medicine, Bristol Heart Institute, University of Bristol, United Kingdom (E.A., I.R.A., H.L.S., F.R., G.M., S.C.S., J.R., V.V.A., O.O.I., S.S., A.O., P.M.).
authors.forename: Federica
authors.initials: F
authors.lastname: Riu
authors.affiliation: Division of Experimental Cardiovascular Medicine, Bristol Heart Institute, University of Bristol, United Kingdom (E.A., I.R.A., H.L.S., F.R., G.M., S.C.S., J.R., V.V.A., O.O.I., S.S., A.O., P.M.).
authors.forename: Giuseppe
authors.initials: G
authors.lastname: Mangialardi
authors.affiliation: Division of Experimental Cardiovascular Medicine, Bristol Heart Institute, University of Bristol, United Kingdom (E.A., I.R.A., H.L.S., F.R., G.M., S.C.S., J.R., V.V.A., O.O.I., S.S., A.O., P.M.).
authors.forename: Sadie C
authors.initials: SC
authors.lastname: Slater
authors.affiliation: Division of Experimental Cardiovascular Medicine, Bristol Heart Institute, University of Bristol, United Kingdom (E.A., I.R.A., H.L.S., F.R., G.M., S.C.S., J.R., V.V.A., O.O.I., S.S., A.O., P.M.).
authors.forename: Jonathan
authors.initials: J
authors.lastname: Rowlinson
authors.affiliation: Division of Experimental Cardiovascular Medicine, Bristol Heart Institute, University of Bristol, United Kingdom (E.A., I.R.A., H.L.S., F.R., G.M., S.C.S., J.R., V.V.A., O.O.I., S.S., A.O., P.M.).
authors.forename: Valeria V
authors.initials: VV
authors.lastname: Alvino
authors.affiliation: Division of Experimental Cardiovascular Medicine, Bristol Heart Institute, University of Bristol, United Kingdom (E.A., I.R.A., H.L.S., F.R., G.M., S.C.S., J.R., V.V.A., O.O.I., S.S., A.O., P.M.).
authors.forename: Oluwasomidotun O
authors.initials: OO
authors.lastname: Idowu
authors.affiliation: Division of Experimental Cardiovascular Medicine, Bristol Heart Institute, University of Bristol, United Kingdom (E.A., I.R.A., H.L.S., F.R., G.M., S.C.S., J.R., V.V.A., O.O.I., S.S., A.O., P.M.).
authors.forename: Stephanie
authors.initials: S
authors.lastname: Soyombo
authors.affiliation: Division of Experimental Cardiovascular Medicine, Bristol Heart Institute, University of Bristol, United Kingdom (E.A., I.R.A., H.L.S., F.R., G.M., S.C.S., J.R., V.V.A., O.O.I., S.S., A.O., P.M.).
authors.forename: Atsuhiko
authors.initials: A
authors.lastname: Oikawa
authors.affiliation: Division of Congenital Heart Surgery, Bristol Heart Institute, University of Bristol, United Kingdom (M.M.S., H.J., M.T.G., M.C.).
authors.forename: Megan M
authors.initials: MM
authors.lastname: Swim
authors.affiliation: School of Physiology and Pharmacology, Bristol Heart Institute, University of Bristol, United Kingdom (C.T.K., H.C., J.C.H., C.H.O.).
authors.forename: Cherrie H T
authors.initials: CH
authors.lastname: Kong
authors.affiliation: School of Physiology and Pharmacology, Bristol Heart Institute, University of Bristol, United Kingdom (C.T.K., H.C., J.C.H., C.H.O.).
authors.forename: Hongwei
authors.initials: H
authors.lastname: Cheng
authors.affiliation: Division of Congenital Heart Surgery, Bristol Heart Institute, University of Bristol, United Kingdom (M.M.S., H.J., M.T.G., M.C.).
authors.forename: Huidong
authors.initials: H
authors.lastname: Jia
authors.affiliation: Division of Congenital Heart Surgery, Bristol Heart Institute, University of Bristol, United Kingdom (M.M.S., H.J., M.T.G., M.C.).
authors.forename: Mohamed T
authors.initials: MT
authors.lastname: Ghorbel
authors.affiliation: School of Physiology and Pharmacology, Bristol Heart Institute, University of Bristol, United Kingdom (C.T.K., H.C., J.C.H., C.H.O.).
authors.forename: Jules C
authors.initials: JC
authors.lastname: Hancox
authors.affiliation: School of Physiology and Pharmacology, Bristol Heart Institute, University of Bristol, United Kingdom (C.T.K., H.C., J.C.H., C.H.O.).
authors.forename: Clive H
authors.initials: CH
authors.lastname: Orchard
authors.affiliation: Division of Cardiac Surgery, Bristol Heart Institute, University of Bristol, United Kingdom (G.A.) Imperial College of London, London, United Kingdom (G.A., C.E.).
authors.forename: Gianni
authors.initials: G
authors.lastname: Angelini
authors.affiliation: Vascular Pathology and Regeneration, Bristol Heart Institute, University of Bristol, United Kingdom (C.E.) Imperial College of London, London, United Kingdom (G.A., C.E.).
authors.forename: Costanza
authors.initials: C
authors.lastname: Emanueli
authors.affiliation: Division of Congenital Heart Surgery, Bristol Heart Institute, University of Bristol, United Kingdom (M.M.S., H.J., M.T.G., M.C.).
authors.forename: Massimo
authors.initials: M
authors.lastname: Caputo
authors.affiliation: Division of Experimental Cardiovascular Medicine, Bristol Heart Institute, University of Bristol, United Kingdom (E.A., I.R.A., H.L.S., F.R., G.M., S.C.S., J.R., V.V.A., O.O.I., S.S., A.O., P.M.).
authors.forename: Paolo
authors.initials: P
authors.lastname: Madeddu
mesh_terms: D003470:Culture Media; D004797:Enzyme-Linked Immunosorbent Assay; D005434:Flow Cytometry; D005455:Fluorescent Antibody Technique; D020869:Gene Expression Profiling; D006330:Heart Defects, Congenital / Q000601:surgery*; D006801:Humans; D007223:Infant; D007231:Infant, Newborn; D020286:Pericytes / Q000166:cytology* / Q000502:physiology; D060888:Real-Time Polymerase Chain Reaction; D013234:Stem Cells / Q000166:cytology / Q000502:physiology; D023822:Tissue Engineering / Q000379:methods*; D016378:Tissue Transplantation / Q000379:methods
chemical_list: D003470:Culture Media
keywords: cells; congenital heart defects; grafting; myocardium; pediatrics