TY - JOUR

T1 - Endothelial and smooth muscle cells derived from human cardiac explants demonstrate angiogenic potential and suitable for design of cell-containing vascular grafts

AU - Zakharova, I. S.

AU - Zhiven', M. K.

AU - Saaya, Sh B.

AU - Shevchenko, A. I.

AU - Smirnova, A. M.

AU - Strunov, A.

AU - Karpenko, A. A.

AU - Pokushalov, E. A.

AU - Ivanova, L. N.

AU - Makarevich, P. I.

AU - Parfyonova, Y. V.

AU - Aboian, E.

AU - Zakian, S. M.

N1 - Publisher Copyright: © 2017 The Author(s).

PY - 2017/3/3

Y1 - 2017/3/3

N2 - Background: 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. Methods: 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. Results: 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. Conclusions: 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.

AB - Background: 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. Methods: 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. Results: 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. Conclusions: 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.

KW - Chitosan

KW - Endothelial cells

KW - Human cardiac explant

KW - Polycaprolactone

KW - Smooth muscle cells

KW - Tissue-engineered vascular grafts

KW - Neovascularization, Physiologic/drug effects

KW - Cell Separation

KW - Humans

KW - Cells, Cultured

KW - Blood Vessel Prosthesis

KW - Cell Survival/drug effects

KW - Mice, SCID

KW - Prosthesis Design

KW - Endothelial Cells/cytology

KW - Animals

KW - Chitosan/pharmacology

KW - Platelet Endothelial Cell Adhesion Molecule-1/metabolism

KW - Weibel-Palade Bodies/metabolism

KW - Polyesters/pharmacology

KW - Myocytes, Smooth Muscle/cytology

KW - Cell Differentiation

KW - Cell Proliferation/drug effects

KW - Myocardium/cytology

KW - TRANSPLANTATION

KW - IN-VITRO

KW - CHITOSAN-POLYCAPROLACTONE BLENDS

KW - PHENOTYPIC HETEROGENEITY

KW - MURINE HEART

KW - PROGENITOR CELLS

KW - STEM-CELLS

KW - CULTURE

KW - ISCHEMIC CARDIOMYOPATHY

KW - GROWTH-FACTOR

UR - http://www.scopus.com/inward/record.url?scp=85014420818&partnerID=8YFLogxK

U2 - 10.1186/s12967-017-1156-1

DO - 10.1186/s12967-017-1156-1

M3 - Article

C2 - 28257636

AN - SCOPUS:85014420818

VL - 15

SP - 54

JO - Journal of Translational Medicine

JF - Journal of Translational Medicine

SN - 1479-5876

IS - 1

M1 - 54

ER -