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

Published: April 30, 2023

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) 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 c…

Representative 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 microscope image of the SMCs seeded on the ECIS plate; magnification 10x. (B) Resistance curves measured by ECIS after seeding of co…

Divulgaciones

The authors have nothing to disclose.

Materials

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

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Measuring SMC Contraction In Vitro: A Non-Invasive Method to Evaluate Smooth Muscle Cells Contraction using Electric Cell-Substrate Impedance Sensing. J. Vis. Exp. (Pending Publication), e21214, doi: (2023).

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