Establishment and Application of the Contraction Assay System Based on 3-dimension Culture of Airway Smooth Muscle Cells

Abstract: Airway smooth muscle cells (ASMCs) have been regarded as the main effector cells responsible for bronchoconstriction and increased respiratory resistance in asthma. The detections of the contractile responses of ASMCs provide important experimental evidences for investigation of the pathophysiological mechanisms of asthma， evaluation and development of new bronchodilator drugs. The routine studies to measure the tension changes of excised tissue and morphological changes of monolayer cultured ASMCs were frequently limited by the intermittent supply of viable samples and measurement error. In this study， we established a gel contraction assay， which employed 3-dimension cultured ASMCs in collagen I substrate. Collagen gels were released from the wells in which they were fabricated， and the decreases in gel surface area caused by cell shortening at different time points were recorded to estimate ASMCs contraction. The results showed that gel area decreased significantly to the stimuli of 0.1， 1， 10 mmol/L of acetylcholine (Ach) in a dose-dependent manner. However， myosin ATP inhibitor BDM (butanedione monoxime) could inhibit the Ach-induced contractile response of ASMCs， which demonstrated the high reproducibility and accuracy of this assay.

CSTR: 32200.14.cjcb.2015.02.0011