{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,4]],"date-time":"2026-10-04T18:06:48Z","timestamp":1791137208213,"version":"4.1.0"},"reference-count":58,"publisher":"Frontiers Media SA","license":[{"start":{"date-parts":[[2023,6,20]],"date-time":"2023-06-20T00:00:00Z","timestamp":1687219200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":["frontiersin.org"],"crossmark-restriction":true},"short-container-title":["Front. Cardiovasc. Med."],"abstract":"Background<\/jats:title>Calcific aortic valve stenosis (AVS) is defined by pathological changes in the aortic valve (AV) and their predominant cell types: valvular interstitial (VICs) and endothelial cells (VECs). Understanding the cellular and molecular mechanisms of this disease is a prerequisite to identify potential pharmacological treatment strategies. In this study, we present a unique aortic valve cell isolation technique to acquire specific human and porcine cell populations and compared VICs and VECs of these species with each other for the first time.<\/jats:p><\/jats:sec>Methods<\/jats:title>AV cells were isolated from tissue obtained from human patients undergoing surgical aortic valve replacement (SAVR) or from porcine hearts. Functional analysis and in vitro<\/jats:italic> experiments revealed that endothelial-to-mesenchymal transition (EndMT) can be induced in hVECs, leading to a significant increase in mesenchymal markers. In vitro<\/jats:italic> calcification experiments of VICs demonstrated pronounced expression of calcification markers and visible calcific deposits in Alizarin Red staining in both species after incubation with pro-calcific media.<\/jats:p><\/jats:sec>Results<\/jats:title>Cells isolated from patient-derived AVs showed mesenchymal and endothelial-specific gene signatures (VIC and VEC, respectively). For instance, von Willebrand factor (vWF<\/jats:italic>) and platelet endothelial adhesion molecule-1 (PECAM1<\/jats:italic>) were upregulated in VECs, while the myofibroblastic markers alpha-smooth muscle actin (\u03b1-SMA<\/jats:italic>) and vimentin (VIM<\/jats:italic>) were downregulated in VECs compared to VICs. Analysis of cell function by migration revealed that VECs are more migratory than VICs. Induction of EndMT in vitro<\/jats:italic> in VECs displayed increased expression of EndMT markers and decreased expression of endothelial markers, confirming their mesenchymal transdifferentiation ability. In vitro<\/jats:italic> calcification of VICs revealed upregulation of alkaline phosphatase (ALPL<\/jats:italic>), a hallmark of calcification. In addition, other calcification-related genes such as osteocalcin (BGLAP<\/jats:italic>) and runt-related factor 2 (RUNX2<\/jats:italic>) were upregulated. Alizarin red staining of calcified cells provided a further layer of confirmation that the isolated cells were VICs with osteoblastic differentiation capacity.<\/jats:p><\/jats:sec>Conclusion<\/jats:title>This study aims to take a first step towards standardizing a reproducible isolation technique for specific human and porcine VEC and VIC populations. A comparison of human and porcine aortic valve cells demonstrated that porcine cells may serve as an alternative cellular model system in settings where human tissue is difficult to obtain.<\/jats:p><\/jats:sec>","DOI":"10.3389\/fcvm.2023.1151028","type":"journal-article","created":{"date-parts":[[2023,6,20]],"date-time":"2023-06-20T06:37:25Z","timestamp":1687243045000},"update-policy":"https:\/\/doi.org\/10.3389\/crossmark-policy","source":"Crossref","is-referenced-by-count":19,"title":["Human and porcine aortic valve endothelial and interstitial cell isolation and characterization"],"prefix":"10.3389","volume":"10","author":[{"given":"D.","family":"Nehl","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"P. 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