ObjectiveTo observe the biological characteristics of valvular interstitial cells in calcific aortic valve disease (CAVD), so as to lay a foundation for future study.MethodsTissue culture inoculation and immunomagnetic bead separation method were used to isolate the valvular interstitial cells from the normal aortic valves and CAVD valves. The morphological and behavioral characteristics of the isolated valvular interstitial cells were observed. Immunocytochemistry and flow cytometry analysis were employed to determine cellular immunophenotype.ResultsCompared to normal valvular interstitial cells, CAVD valvular interstitial cells displayed a myofibroblast- and osteoblast-like morphology. When the cell density reached a certain level, they spontaneously retracted from the neighboring areas and grouped into aggregates, forming calcific nodules. Furthermore, CAVD valvular interstitial cells cultured in vitro were positive for myofibroblast marker α-SMA and osteoblast marker alkaline phosphatase.ConclusionBiological characteristic change of CAVD valvular interstitial cells might be the major reason for the thickening, calcification, and commissural fusion in CAVD valvular samples.