Fabrication procedure for SMC sheet containing layered elastic fibers. (a) Vascular SMCs were seeded on a cell culture disk to make the first SMC layer. Beginning 24 hours after seeding, SMCs were exposed to periodic hydrostatic pressure (PHP) for 24 hours. This PHP promoted SMC-derived fibronectin fibrillogenesis (illustrated as green color), which enabled the SMCs of the second layer to attach to the first layer. This procedure was repeated up to ten times to form a ten-layered SMC sheet. AP: atmospheric pressure. (b) A section of our ten-layered SMC sheet was stained with the Elastica van Gieson stain and the Masson trichrome stain to reveal the elastic fibers (deep purple color, upper panel) and collagens (blue color, lower panel), respectively. Scale bars: 100 μm. (c) Our SMC sheet was able to be stretched with a DMT560 tissue puller (Danish Myo Technology, Aarhus N, Denmark). Stretched and unstretched SMC sheets are shown.