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…
Divulgazioni
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 |
Tags
Citazione di questo articolo
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).