[Real-time Measurement of Cell Contractile Force During Activation of Human Hepatic Stellate Cell Line LX-2]

The contractile force of hepatic stellate cells plays a very important role in liver damage, hepatitis and fibrosis. In this paper, a method based on polydimethylsiloxane (PDMS) thin micropillar arrays is proposed to measure the contractile force of human hepatic stellate cell line LX-2, which enables dynamic measurement of the subcellular distribution of force magnitude and direction. First, thin micropillar arrays on glass bottom dish were fabricated using two-step casting process in order to meet the working distance requirement of 100× objective lens. After hydrophilic treatment and protein imprint, cells were seeded on the micropillar arrays. LX-2 cells, which were quiesced by growth in serum-free medium, were activated by adding fetal bovine serum (FBS). The deflections of the micropillars were achieved by image processing technique, and then the contractile force of cells exerted on the micropillars was calculated according to mechanical simulation results, and was analyzed under both quiescent and activated conditions. The experimental results show that the average traction force of quiescent cells is about 20 nN, while the contractile force of activated cells increased to 110 nN upon adding FBS. This method can quantify the contractile force of LX-2 cell on subcellular scale in both quiescent and activated states, which may benefit pathology study and drug screen for chronic liver diseases resulted from liver fibrosis.