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An immunofluorescence assay for extracellular matrix components highlights the role of epithelial cells in producing a stable, fibrillar extracellular matrix
- Source :
- Biology Open, Vol 6, Iss 10, Pp 1423-1433 (2017), Biology Open
- Publication Year :
- 2017
- Publisher :
- The Company of Biologists, 2017.
-
Abstract
- Activated fibroblasts are considered major drivers of fibrotic disease progression through the production of excessive extracellular matrix (ECM) in response to signals from damaged epithelial and inflammatory cells. Nevertheless, epithelial cells are capable of expressing components of the ECM, cross-linking enzymes that increase its stability and are sensitive to factors involved in the early stages of fibrosis. We therefore wanted to test the hypothesis that epithelial cells can deposit ECM in response to stimulation in a comparable manner to fibroblasts. We performed immunofluorescence analysis of components of stable, mature extracellular matrix produced by primary human renal proximal tubular epithelial cells and renal fibroblasts in response to cytokine stimulation. Whilst fibroblasts produced a higher basal level of extracellular matrix components, epithelial cells were able to deposit significant levels of fibronectin, collagen I, III and IV in response to cytokine stimulation. In response to hypoxia, epithelial cells showed an increase in collagen IV deposition but not in response to the acute stress stimuli aristolochic acid or hydrogen peroxide. When epithelial cells were in co-culture with fibroblasts we observed significant increases in the level of matrix deposition which could be reduced by transforming growth factor beta (TGF-β) blockade. Our results highlight the role of epithelial cells acting as efficient producers of stable extracellular matrix which could contribute to renal tubule thickening in fibrosis.<br />Summary: Using an immunofluorescence extracellular matrix, we show that epithelial cells can deposit extracellular matrix components in a comparable manner to fibroblasts highlighting their role as potential targets in fibrotic disease.
- Subjects :
- 0301 basic medicine
Pathology
medicine.medical_specialty
Tissue transglutaminase
QH301-705.5
Science
030232 urology & nephrology
Matrix (biology)
Kidney
Transforming growth factor beta (TGF-β)
General Biochemistry, Genetics and Molecular Biology
Extracellular matrix
03 medical and health sciences
0302 clinical medicine
Fibrosis
medicine
Biology (General)
Fibroblast
Fibronectin
Epithelial polarity
biology
Transforming growth factor beta
medicine.disease
Transglutaminase
Cell biology
Epithelial cell
030104 developmental biology
medicine.anatomical_structure
biology.protein
Collagen
General Agricultural and Biological Sciences
Research Article
Subjects
Details
- Language :
- English
- ISSN :
- 20466390
- Volume :
- 6
- Issue :
- 10
- Database :
- OpenAIRE
- Journal :
- Biology Open
- Accession number :
- edsair.doi.dedup.....651b12d029df73c8d635b6562e4c0ee6