A dynamic real-time method for monitoring epithelial barrier function in vitro

Anal Biochem. 2012 Jun 15;425(2):96-103. doi: 10.1016/j.ab.2012.03.010. Epub 2012 Mar 23.


The intestinal epithelium functions as a physical barrier against the harmful environment of the lumen, which usually becomes impaired in the presence of intestinal diseases. In this work, we introduce an electronic impedance-based analysis using a real-time xCELLigence system to record the dynamic processes of ethanol-induced intestinal barrier dysfunction. In terms of analyzing morphological alterations in the paracellular junction complex and the organization of pericellular F-actin, this novel, real-time, cell-based technology shows considerable correlations with the standard transepithelial electrical resistance endpoint assay. In addition, monitoring barrier functions in real time allows unbiased screening and characterization of biochemical agents in the lumen that affect epithelial integrity. This functional assay further identifies the in vitro roles of the inducible nitric oxide synthase inhibitor, epithelial growth factor, tyrosine kinases, and phosphatases in regulating epithelial barrier function in response to ethanol administration. Taken together, our findings suggest that this novel, real-time, high-throughput method offers a promising tool for monitoring epithelial barrier functions in situations with more physiological relevance.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Actins / metabolism
  • Biochemistry / methods*
  • Caco-2 Cells
  • Cell Membrane Permeability / drug effects
  • Electric Impedance
  • Electrochemical Techniques
  • Ethanol / toxicity
  • Humans
  • Intestinal Mucosa / drug effects*
  • Intestinal Mucosa / metabolism
  • Nitric Oxide Synthase Type II / antagonists & inhibitors
  • Nitric Oxide Synthase Type II / metabolism
  • Protein Kinase Inhibitors / pharmacology


  • Actins
  • Protein Kinase Inhibitors
  • Ethanol
  • Nitric Oxide Synthase Type II