Measuring SMC Contraction In Vitro: A Non-Invasive Method to Evaluate Smooth Muscle Cells Contraction using Electric Cell-Substrate Impedance Sensing

Count the cells using an automated cell counter, and seed the SMCs in triplicate at a seeding density of 30,000 cells per well, in 200 microliters of SMC medium in the gelatin-coated ECIS plate. Place the plate with the SMCs into the ECIS 96-well holder in the cell culture incubator.

Double-click on the "ECIS Applied Biophysics software" to open the program, and press the "Setup" button. Check if all the electrodes have contact with the holder, in the left lower panel labeled "Well Configuration." If the electrodes are not properly connected, adjust the plate in the holder before starting the measurement.

Now, select the plate type in the same panel by clicking "Array Type." Next, prepare two pipettes set at 2-microliter and 150-microliter volume. Before starting the stimulation, press "Mark" in the software, and place a comment.

To stimulate the cells, remove the lid, and place it on a sterile surface inside the incubator. Then, induce SMC contraction by pipetting 2 microliters of the ionomycin working solution into each well as quickly as possible.

Once all the cells have been stimulated, mix the medium in the wells using the second pipette.