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Robust generation of quiescent porcine valvular interstitial cell cultures
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نویسنده
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porras a.m. ,van engeland n.c.a. ,marchbanks e. ,mccormack a. ,bouten c.v.c. ,yacoub m.h. ,latif n. ,masters k.s.
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منبع
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journal of the american heart association - 2017 - دوره : 6 - شماره : 3
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چکیده
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Background--valvular interstitial cells (vics) in the healthy aortic valve leaflet exhibit a quiescent phenotype,with < 5% of vics exhibiting an activated phenotype. yet,in vitro culture of vics on tissue culture polystyrene surfaces in standard growth medium results in rapid transformation to an activated phenotype in > 90% of cells. the inability to preserve a healthy vic phenotype during in vitro studies has hampered the elucidation of mechanisms involved in calcific aortic valve disease. this study describes the generation of quiescent populations of porcine vics in 2-dimensional in vitro culture and their utility in studying valve pathobiology. methods and results--within 4 days of isolation from fresh porcine hearts,vics cultured in standard growth conditions were predominantly myofibroblastic (activated vics). this myofibroblastic phenotype was partially reversed within 4 days,and fully reversed within 9 days,following application of a combination of a fibroblast media formulation with culture on collagen coatings. specifically,culture in this combination significantly reduced several markers of vic activation,including proliferation,apoptosis,a-smooth muscle actin expression,and matrix production,relative to standard growth conditions. moreover,vics raised in a fibroblast media formulation with culture on collagen coatings exhibited dramatically increased sensitivity to treatment with transforming growth factor b1,a known pathological stimulus,compared with vics raised in either standard culture or medium with a fibroblast media formulation. conclusions--the approach using a fibroblast media formulation with culture on collagen coatings generates quiescent vics that more accurately mimic a healthy vic population and thus has the potential to transform the study of the mechanisms of vic activation and dysfunction involved in the early stages of calcific aortic valve disease. © 2017 the authors.
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کلیدواژه
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Aortic valve; Calcific aortic valve disease; Differentiation; Myofibroblasts; Quiescence; Valvular interstitial cell
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آدرس
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department of biomedical engineering,university of wisconsin-madison,madison,wi, United States, department of biomedical engineering,eindhoven university of technology,eindhoven, Netherlands, department of chemical and biological engineering,colorado school of mines,golden,co, United States, tissue engineering,magdi yacoub institute,imperial college,london, United Kingdom, department of biomedical engineering,eindhoven university of technology,eindhoven, Netherlands, tissue engineering,magdi yacoub institute,imperial college,london, United Kingdom, tissue engineering,magdi yacoub institute,imperial college,london, United Kingdom, department of biomedical engineering,university of wisconsin-madison,madison,wi, United States
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Authors
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