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

Abstract

Source: Bogunovic, N. et al., Isolation of Primary Patient-specific Aortic Smooth Muscle Cells and Semiquantitative Real-time Contraction Measurements In Vitro. J. Vis. Exp. (2022).

This video describes the measurement of aortic smooth muscle cell (SMC) contraction using electrical cell-substrate impedance sensing, or ECIS. ECIS measures the locomotion of cells and is a reliable method for measuring contractions.

Protocol

All procedures involving human participants have been performed in compliance with the institutional, national, and international guidelines for human welfare and have been reviewed by the local institutional review board

1. Measuring contraction (example SMCs)

1. Prepare a 96w10e ECIS array (see Figure 1A for a representative image of the array with magnified electrodes and cells seeded on the electrodes).
   NOTE: Perform the following steps under a sterile tissue culture laminar flow hood.
   1. Coat a sterile 96w10e array with 200 µL of 10 mM L-cysteine per well for 30 min at room temperature.
   ....

Results

Figure 1: ECIS plate setup and graphic representation of aortic SMC contraction. (A) Left; a 96-well ECIS plate. Middle; magnification of one well of the ECIS plate, showing the ten electrodes at the bottom of the well. Right; light micro.......

Materials

List of materials used in this article

Name	Company	Catalog Number	Comments
96-well Array	Applied Biophysics	96W10idf PET	Array used to measure contraction in the ECIS setup
Human Vascular Smooth Muscle Cell Basal Medium (formerly ''Medium 231'')	Gibco	M231500	Medium used to culture smooth muscle cells
Dimethyl sulfoxide	Sigma-Aldrich	472301	Solution used to dilute ionomycin
Invitrogen countess II	Thermo Fisher Scientific	AMQAX1000	Automated cell counter
Ionomycin calcium salt from Streptomyces conglobatus	Sigma-Aldrich	I0634-1MG	Compound used for contraction stimulation
ZTheta	Applied Biophysics	ZTheta	ECIS instrument used for contraction measurements