Objective To investigate the influence of three extracellular matrix (ECM) proteins, namely, laminin (LN), collagen type I (Col I), fibronectin (FN) on the morphology and contractility changes in airway smooth muscle cells (ASMCs) induced by platelet-derived growth factor-BB (PDGF-BB). Methods ASMCs were seeded on the culture dish coated with LN, Col I, or FN, respectively, and divided into two groups to be cultured either in the absence or presence of PDGF-BB (10 mg/L) for 0~5 d. Subsequently, cell morphology was examined by the optical microscopy and quantified as the ratio of cell width to length, and the KCl/histamine-induced contractile responses of the cell were measured by optical magnetic twisting cytometry (OMTC). Results ASMCs cultured in the presence of PDGF-BB generally appeared in longer and thinner cell shapes, namely, a smaller ratio of cell width to length, but the cell width/length ratio for ASMCs adhered on LN was relatively bigger than that on Col I or FN. In the absence of PDGF-BB, contractility of ASMCs to KCl increased with the duration of culture, which was independent of the ECM proteins. In contrast, in the presence of PDGF-BB, contractility of ASMCs to KCl or histamine decreased in all situations, but degree of the decrease was smaller for ASMCs adhered on LN than those on Col I or FN. Conclusions The morphology and contractility changes in ASMCs induced by PDGF-BB are influenced by ECM proteins on which cells are grown. For ASMCs adhered on LN, the morphology and contractility changes are relatively smaller than those on Col I and FN. The differential effect of ECM proteins on PDGF-BB induced changes in morphology and contractility of ASMCs is important to fully understand the interactions between ECM proteins, inflammatory factors, ASMCs, and their relation to the pathophysiological mechanism of asthma.