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J. Cardiovasc. Dev. Dis., Volume 1, Issue 2 (September 2014), Pages 146-176

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Research

Open AccessArticle Characterization of Dermal Fibroblasts as a Cell Source for Pediatric Tissue Engineered Heart Valves
J. Cardiovasc. Dev. Dis. 2014, 1(2), 146-162; doi:10.3390/jcdd1020146
Received: 17 June 2014 / Revised: 29 July 2014 / Accepted: 20 August 2014 / Published: 26 August 2014
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Abstract
There is continued debate regarding the appropriate cell type to replace valvular interstitial cells (VICs) in tissue engineered heart valves (TEHVs), particularly for pediatric patients. In this work, neonatal human dermal fibroblasts (nhDFFs) were compared to human pediatric VICs (hpVICs), based on their
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There is continued debate regarding the appropriate cell type to replace valvular interstitial cells (VICs) in tissue engineered heart valves (TEHVs), particularly for pediatric patients. In this work, neonatal human dermal fibroblasts (nhDFFs) were compared to human pediatric VICs (hpVICs), based on their phenotypic and gene expression characteristics when cultured on collagen type I, fibronectin, fibrin, and tissue culture polystyrene (TCP) substrates. Similar confluency was achieved over the culture period on collagen and fibronectin between both cell types, although nhDFFs tended to reach lower confluence on collagen than on any other substrate. Morphologically, hpVICs tended to spread and form multiple extensions, while nhDFFs remained homogenously spindle-shaped on all substrates. PCR results indicated that fibroblasts did not differ significantly from VICs in gene expression when cultured on fibrin, whereas on collagen type I and fibronectin they showed increased α-SMA, xylosyltransferase I, and collagen type I expression (p < 0.05). However, protein expression of these targets, analyzed by immunocytochemistry and Western blotting, was not significantly different between cell types. These results suggest that nhDFFs express similar matrix production and remodeling genes as hpVICs, and the choice of substrate for TEHV construction can affect the growth and expression profile of nhDFFs as compared to native hpVICs. Full article
(This article belongs to the Special Issue Semilunar Valve Development and Disease)
Figures

Open AccessArticle Sox9- and Scleraxis-Cre Lineage Fate Mapping in Aortic and Mitral Valve Structures
J. Cardiovasc. Dev. Dis. 2014, 1(2), 163-176; doi:10.3390/jcdd1020163
Received: 26 August 2014 / Revised: 16 September 2014 / Accepted: 17 September 2014 / Published: 23 September 2014
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Abstract
Heart valves are complex structures composed of a heterogeneous population of valve interstitial cells (VICs), an overlying endothelium and highly organized layers of extracellular matrix. Alterations in valve homeostasis are characteristic of dysfunction and disease, however the mechanisms that initiate and promote valve
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Heart valves are complex structures composed of a heterogeneous population of valve interstitial cells (VICs), an overlying endothelium and highly organized layers of extracellular matrix. Alterations in valve homeostasis are characteristic of dysfunction and disease, however the mechanisms that initiate and promote valve pathology are poorly understood. Advancements have been largely hindered by the limited availability of tools for gene targeting in heart valve structures during embryogenesis and after birth. We have previously shown that the transcription factors Sox9 and Scleraxis (Scx) are required for heart valve formation and in this study we describe the recombination patterns of Sox9- and Scx-Cre lines at differential time points in aortic and mitral valve structures. In ScxCre; ROSA26GFP mice, recombination is undetected in valve endothelial cells (VECs) and low in VICs during embryogenesis. However, recombination increases in VICs from post natal stages and by 4 weeks side-specific patterns are observed. Using the inducible Sox9CreERT2 system, we observe recombination in VECs and VICs in the embryo, and high levels are maintained through post natal and juvenile stages. These Cre-drivers provide the field with new tools for gene targeting in valve cell lineages during differential stages of embryonic and post natal maturation and maintenance. Full article
(This article belongs to the Special Issue Semilunar Valve Development and Disease)

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