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Endothelial and smooth muscle cells derived from human cardiac explants demonstrate angiogenic potential and suitable for design of cell-containing vascular grafts. / Zakharova, I. S.; Zhiven', M. K.; Saaya, Sh B. et al.
In: Journal of Translational Medicine, Vol. 15, No. 1, 54, 03.03.2017, p. 54.Research output: Contribution to journal › Article › peer-review
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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 -
ID: 8754350