title: Endothelial and smooth muscle cells derived from human cardiac explants demonstrate angiogenic potential and suitable for design of cell-containing vascular grafts.
abstract: Endothelial and smooth muscle cells are considered promising resources for regenerative medicine and cell replacement therapy. It has been shown that both types of cells are heterogeneous depending on the type of vessels and organs in which they are located. Therefore, isolation of endothelial and smooth muscle cells from tissues relevant to the area of research is necessary for the adequate study of specific pathologies. However, sources of specialized human endothelial and smooth muscle cells are limited, and the search for new sources is still relevant. The main goal of our study is to demonstrate that functional endothelial and smooth muscle cells can be obtained from an available source-post-surgically discarded cardiac tissue from the right atrial appendage and right ventricular myocardium.
  
  
  
  Heterogeneous primary cell cultures were enzymatically isolated from cardiac explants and then grown in specific endothelial and smooth muscle growth media on collagen IV-coated surfaces. The population of endothelial cells was further enriched by immunomagnetic sorting for CD31, and the culture thus obtained was characterized by immunocytochemistry, ultrastructural analysis and in vitro functional tests. The angiogenic potency of the cells was examined by injecting them, along with Matrigel, into immunodeficient mice. Cells were also seeded on characterized polycaprolactone/chitosan membranes with subsequent analysis of cell proliferation and function.
  
  
  
  Endothelial cells isolated from cardiac explants expressed CD31, VE-cadherin and VEGFR2 and showed typical properties, namely, cytoplasmic Weibel-Palade bodies, metabolism of acetylated low-density lipoproteins, formation of capillary-like structures in Matrigel, and production of extracellular matrix and angiogenic cytokines. Isolated smooth muscle cells expressed extracellular matrix components as well as α-actin and myosin heavy chain. Vascular cells derived from cardiac explants demonstrated the ability to stimulate angiogenesis in vivo. Endothelial cells proliferated most effectively on membranes made of polycaprolactone and chitosan blended in a 25:75 ratio, neutralized by a mixture of alkaline and ethanol. Endothelial and smooth muscle cells retained their functional properties when seeded on the blended membranes.
  
  
  
  We established endothelial and smooth muscle cell cultures from human right atrial appendage and right ventricle post-operative explants. The isolated cells revealed angiogenic potential and may be a promising source of patient-specific cells for regenerative medicine.
authors.affiliation: The Federal Research Center Institute of Cytology And Genetics, The Siberian Branch of the Russian Academy of Sciences, Novosibirsk, Russian Federation. zakharova@bionet.nsc.ru.|Institute of Chemical Biology and Fundamental Medicine, The Siberian Branch of the Russian Academy of Sciences, Novosibirsk, Russian Federation. zakharova@bionet.nsc.ru.|Siberian Federal Biomedical Research Center, Ministry of Health Care of Russian Federation, Novosibirsk, Russian Federation. zakharova@bionet.nsc.ru.
authors.forename: I S
authors.initials: IS
authors.lastname: Zakharova
authors.affiliation: The Federal Research Center Institute of Cytology And Genetics, The Siberian Branch of the Russian Academy of Sciences, Novosibirsk, Russian Federation.|Institute of Chemical Biology and Fundamental Medicine, The Siberian Branch of the Russian Academy of Sciences, Novosibirsk, Russian Federation.|Siberian Federal Biomedical Research Center, Ministry of Health Care of Russian Federation, Novosibirsk, Russian Federation.
authors.forename: M K
authors.initials: MK
authors.lastname: Zhiven'
authors.affiliation: Siberian Federal Biomedical Research Center, Ministry of Health Care of Russian Federation, Novosibirsk, Russian Federation.
authors.forename: Sh B
authors.initials: SB
authors.lastname: Saaya
authors.affiliation: The Federal Research Center Institute of Cytology And Genetics, The Siberian Branch of the Russian Academy of Sciences, Novosibirsk, Russian Federation.|Institute of Chemical Biology and Fundamental Medicine, The Siberian Branch of the Russian Academy of Sciences, Novosibirsk, Russian Federation.|Siberian Federal Biomedical Research Center, Ministry of Health Care of Russian Federation, Novosibirsk, Russian Federation.|Novosibirsk State University, Novosibirsk, Russian Federation.
authors.forename: A I
authors.initials: AI
authors.lastname: Shevchenko
authors.affiliation: The Federal Research Center Institute of Cytology And Genetics, The Siberian Branch of the Russian Academy of Sciences, Novosibirsk, Russian Federation.|Siberian Federal Biomedical Research Center, Ministry of Health Care of Russian Federation, Novosibirsk, Russian Federation.|Novosibirsk State University, Novosibirsk, Russian Federation.
authors.forename: A M
authors.initials: AM
authors.lastname: Smirnova
authors.affiliation: The Federal Research Center Institute of Cytology And Genetics, The Siberian Branch of the Russian Academy of Sciences, Novosibirsk, Russian Federation.
authors.forename: A
authors.initials: A
authors.lastname: Strunov
authors.affiliation: Siberian Federal Biomedical Research Center, Ministry of Health Care of Russian Federation, Novosibirsk, Russian Federation.
authors.forename: A A
authors.initials: AA
authors.lastname: Karpenko
authors.affiliation: Siberian Federal Biomedical Research Center, Ministry of Health Care of Russian Federation, Novosibirsk, Russian Federation.
authors.forename: E A
authors.initials: EA
authors.lastname: Pokushalov
authors.affiliation: The Federal Research Center Institute of Cytology And Genetics, The Siberian Branch of the Russian Academy of Sciences, Novosibirsk, Russian Federation.|Novosibirsk State University, Novosibirsk, Russian Federation.
authors.forename: L N
authors.initials: LN
authors.lastname: Ivanova
authors.affiliation: Laboratory of Angiogenesis, Russian Cardiology Research and Production Complex, Moscow, Russian Federation.|Laboratory of gene and cell therapy, Institute of regenerative medicine, Lomonosov Moscow State University, Moscow, Russian Federation.
authors.forename: P I
authors.initials: PI
authors.lastname: Makarevich
authors.affiliation: Laboratory of Angiogenesis, Russian Cardiology Research and Production Complex, Moscow, Russian Federation.|Faculty of Medicine, Lomonosov Moscow State University, Moscow, Russian Federation.
authors.forename: Y V
authors.initials: YV
authors.lastname: Parfyonova
authors.affiliation: Division of Vascular Surgery, Palo Alto Medical Foundation, Burlingame, USA.
authors.forename: E
authors.initials: E
authors.lastname: Aboian
authors.affiliation: The Federal Research Center Institute of Cytology And Genetics, The Siberian Branch of the Russian Academy of Sciences, Novosibirsk, Russian Federation.|Institute of Chemical Biology and Fundamental Medicine, The Siberian Branch of the Russian Academy of Sciences, Novosibirsk, Russian Federation.|Siberian Federal Biomedical Research Center, Ministry of Health Care of Russian Federation, Novosibirsk, Russian Federation.|Novosibirsk State University, Novosibirsk, Russian Federation.
authors.forename: S M
authors.initials: SM
authors.lastname: Zakian
mesh_terms: D000818:Animals; D001807:Blood Vessel Prosthesis; D002454:Cell Differentiation; D049109:Cell Proliferation / Q000187:drug effects; D002469:Cell Separation; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D048271:Chitosan / Q000494:pharmacology; D042783:Endothelial Cells / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism / Q000648:ultrastructure; D006801:Humans; D016513:Mice, SCID; D009206:Myocardium / Q000166:cytology*; D032389:Myocytes, Smooth Muscle / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism / Q000648:ultrastructure; D018919:Neovascularization, Physiologic / Q000187:drug effects; D019408:Platelet Endothelial Cell Adhesion Molecule-1 / Q000378:metabolism; D011091:Polyesters / Q000494:pharmacology; D011474:Prosthesis Design; D020302:Weibel-Palade Bodies / Q000378:metabolism
chemical_list: D019408:Platelet Endothelial Cell Adhesion Molecule-1; D011091:Polyesters; C016240:polycaprolactone; D048271:Chitosan
keywords: Chitosan; Endothelial cells; Human cardiac explant; Polycaprolactone; Smooth muscle cells; Tissue-engineered vascular grafts