Quiescent hepatic stellate cells (qHSCs) possess processes and surround the sinusoids, the capillaries of the liver. We have suggested that HSCs regulate hepatic blood flow under physiological conditions. However, there is no direct evidence whether qHSCs contract. In this study, we established a new method to evaluate the contraction of cells and used this method for studying an effect of receptor agonist on the contractile function of qHSCs. qHSCs isolated from mice were seeded on collagen gel with fluorescent beads, so that beads were attracted to contracting qHSCs. By measuring the distance of the beads movement, we evaluated the contraction. 10 nM endothelin-1 (ET-1) induced slow contraction of qHSC and 1μM bosentan inhibited this contraction. This suggested ET-1 make qHSCs contract via endothelin receptor. Myosin light chain kinase (MLCK) inhibitor, ML-9 (1 μM) inhibited this contraction. Calcium free condition reduced ET-1-induced contraction but still sustained contraction was remained. This was diminished by combination use with 1 μM H-1152, the Rho kinase inhibitor. These results suggest that ET-1 induces contraction in qHSCs which is mediated by calcium-MLCK system and Rho-kinase system.