Asporin Reduces Adult Aortic Valve Interstitial Cell Mineralization Induced by Osteogenic Media and Wnt Signaling Manipulation In Vitro

Figure 1

Isolation and characterization of avian adult aortic valve interstitial cells (AVICs) in vitro. (a) Light microscopic image of tri-cusps at the basement of adult avian aortic valves (arrowheads). (b) Bright filed image (×10) of primary chick AVICs culture. AVIC cells are characterized with filamentous morphology (shown by arrowheads) after 48-72 hr culture. (c) Bright-field images (×20) of Coomassie brilliant blue-stained AVICs with distinct morphology are taken from cultured avian AVIC cells. (d–f) AVICs were immunostained with α-Sma (green), marking myofibroblasts, and counterstained with DAPI (blue) positive nuclei (×10 (d), ×20 (e), and ×40 (f); marked by arrowheads). (g) Gene expression analyses of AVIC-specific markers and Asporin, comparing with ventricle (VEN) markers, by RT-PCR. Here, increased expressions of Fibronectin, Sm22α, and Asporin are detected in AVICs compared to the VEN tissue, and in contrast, VEN-specific markers (Myh7 and Gata4) have shown increased expression in the isolated VEN tissue compared to AVIC cells. β-Actin was used for equal mRNA input and RT efficiency ().

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