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Intestinal epithelial wound healing assay in an epithelial–mesenchymal co‐culture system
Author(s) -
Seltana Amira,
Basora Nuria,
Beaulieu JeanFrançois
Publication year - 2010
Publication title -
wound repair and regeneration
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.847
H-Index - 109
eISSN - 1524-475X
pISSN - 1067-1927
DOI - 10.1111/j.1524-475x.2009.00554.x
Subject(s) - wound healing , basement membrane , epithelium , mesenchymal stem cell , myofibroblast , microbiology and biotechnology , fibronectin , cell culture , chemistry , biology , pathology , extracellular matrix , immunology , medicine , fibrosis , genetics
Rapid and efficient healing of epithelial damage is critical to the functional integrity of the small intestine. Epithelial repair is a complex process that has largely been studied in cultured epithelium but to a much lesser extent in mucosa. We describe a novel method for the study of wound healing using a co‐culture system that combined an intestinal epithelial Caco‐2/15 cell monolayer cultured on top of human intestinal myofibroblasts, which together formed a basement membrane‐like structure that contained many of the major components found at the epithelial–mesenchymal interface in the human intestine. To investigate the mechanism of restitution, small lesions were generated in epithelial cell monolayers on plastic or in co‐cultures without disturbing the underlying mesenchymal layer. Monitoring of wound healing showed that repair was more efficient in Caco‐2/15–myofibroblast co‐cultures than in Caco‐2/15 monolayers and involved the deposition of basement membrane components. Functional experiments showed that the addition of type I collagen or human fibronectin to the culture medium significantly accelerated wound closure on epithelial cell co‐cultures. This system may provide a new tool to investigate the mechanisms that regulate wound healing in the intestinal epithelium.