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Electric Cell-substrate Impedance Sensing for the Quantification of Endothelial Proliferation, Barrier Function, and Motility
Published on: March 28, 2014
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Measuring Cell Growth and Junction Development in Epithelial Cells Using Electric Cell-Substrate Impedance Sensing
Shaista Anwer1, Katalin Szaszi1,2
1Keenan Research Centre for Biomedical Science of the St. Michael's Hospital, Toronto, Canada.
Bio-Protocol
|March 4, 2021
Summary
Electric Cell-substrate Impedance Sensing (ECIS) measures cell growth and barrier function simultaneously. This study shows ECIS can track how claudin-2 depletion impacts tubular epithelial cell development and tight junction integrity.
Area of Science:
- Biotechnology and Bioengineering
- Cell Biology
- Epithelial Biology
Background:
- Electric Cell-substrate Impedance Sensing (ECIS) is a non-invasive technique for real-time monitoring of cellular functions.
- ECIS quantifies cell attachment, proliferation, migration, and the integrity of epithelial barriers, specifically tight junctions.
- Simultaneous assessment of cell number and barrier function provides valuable insights into cellular processes.
Purpose of the Study:
- To describe and utilize the ECIS method for real-time monitoring of cell growth and tight junction-dependent barrier formation.
- To investigate the impact of claudin-2 depletion on tubular epithelial cell growth and barrier function using ECIS.
- To demonstrate the utility of ECIS in addressing fundamental questions regarding tight junction proteins in cell behavior.
Main Methods:
- Cells were transfected with control or claudin-2-specific siRNA.
- Transfected cells were plated on electrodes for ECIS analysis.
- ECIS measurements of capacitance (cell number) and impedance (barrier function) were recorded over time.
- Data analysis involved ECIS software followed by export to graph software.
Main Results:
- ECIS successfully provided simultaneous, real-time data on tubular epithelial cell growth and barrier function development.
- The study quantified the effects of claudin-2 depletion on cell proliferation and the establishment of tight junction barrier.
- Capacitance changes correlated with cell number, reflecting growth and migration, while impedance changes indicated barrier integrity.
Conclusions:
- ECIS is a powerful tool for real-time, simultaneous assessment of cell growth and tight junction barrier function in epithelial cells.
- Claudin-2 plays a significant role in regulating tubular epithelial cell growth and the formation of functional tight junctions.
- This protocol enables detailed investigation into the contribution of specific tight junction proteins to epithelial cell physiology.
