Heart valves leaflets consists of three layers: ventricularis, fibrosa, and spongiosa. In an aortic valve, the ventricularis faces the ventricular compartment, the fibrosa faces the aorta, and the spongiosa is sandwiched between the two layers. The outermost surfaces on both ventricularis and fibrosa are covered with valve endothelial cells (VECs). Underneath the VECs is a sublayer of valve interstitial cells (VICs). Valvular remodeling often involves paracrine regulation between VECs and VICs, and diseases such as valve calcification can result from improper communication between these cells. VECs are known to respond to hemodynamic stimuli, and studies have shown that VECs cultured under disturbed flow undermines endothelial regulation, resulting in increased tissue calcification. In the current investigation, we specifically examined VIC calcification levels after exposure to conditioned media from VECs cultured under various flow environments.