Characterisation of feline renal cortical fibroblast cultures and their transcriptional response to transforming growth factor β1
Abstract Background Chronic kidney disease (CKD) is common in geriatric cats, and the most prevalent pathology is chronic tubulointerstitial inflammation and fibrosis. The cell type predominantly responsible for the production of extra-cellular matrix in renal fibrosis is the myofibroblast, and fibr...
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| Format: | Article |
| Language: | English |
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BMC
2018-03-01
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| Series: | BMC Veterinary Research |
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| Online Access: | http://link.springer.com/article/10.1186/s12917-018-1387-2 |
| _version_ | 1818503958842310656 |
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| author | J. S. Lawson H. M. Syme C. P. D. Wheeler-Jones J. Elliott |
| author_facet | J. S. Lawson H. M. Syme C. P. D. Wheeler-Jones J. Elliott |
| author_sort | J. S. Lawson |
| collection | DOAJ |
| description | Abstract Background Chronic kidney disease (CKD) is common in geriatric cats, and the most prevalent pathology is chronic tubulointerstitial inflammation and fibrosis. The cell type predominantly responsible for the production of extra-cellular matrix in renal fibrosis is the myofibroblast, and fibroblast to myofibroblast differentiation is probably a crucial event. The cytokine TGF-β1 is reportedly the most important regulator of myofibroblastic differentiation in other species. The aim of this study was to isolate and characterise renal fibroblasts from cadaverous kidney tissue of cats with and without CKD, and to investigate the transcriptional response to TGF-β1. Results Cortical fibroblast cultures were successfully established from the kidney tissue of cats with normal kidney function (FCF) and cats with chronic kidney disease (CKD-FCF). Both cell types expressed the mesenchymal markers vimentin, CD44 and CD29, and were negative for the epithelial marker cytokeratin, mesangial cell marker desmin and endothelial cell marker vWF. Only CKD-FCF expressed VCAM-1, a cell marker associated with inflammation. Incubation with TGF-β1 (0–10 ng/ml) induced a concentration dependent change in cell morphology, and upregulation of myofibroblast marker gene α-SMA expression alongside collagen 1α1, fibronectin, TGF-β1 and CTGF mRNA. These changes were blocked by the TGF-β1 receptor 1 antagonist SB431542 (5 μM). Conclusions FCF and CKD-FCF can be cultured via a simple method and represent a model for the investigation of the progression of fibrosis in feline CKD. The findings of this study suggest TGF-β1 may be involved in fibroblast-myofibroblast transition in feline CKD, as in other species. |
| first_indexed | 2024-12-10T21:30:46Z |
| format | Article |
| id | doaj.art-ad6ae265288b47ffbdeccf6bf45888b9 |
| institution | Directory Open Access Journal |
| issn | 1746-6148 |
| language | English |
| last_indexed | 2024-12-10T21:30:46Z |
| publishDate | 2018-03-01 |
| publisher | BMC |
| record_format | Article |
| series | BMC Veterinary Research |
| spelling | doaj.art-ad6ae265288b47ffbdeccf6bf45888b92022-12-22T01:32:49ZengBMCBMC Veterinary Research1746-61482018-03-0114111110.1186/s12917-018-1387-2Characterisation of feline renal cortical fibroblast cultures and their transcriptional response to transforming growth factor β1J. S. Lawson0H. M. Syme1C. P. D. Wheeler-Jones2J. Elliott3Comparative Biomedical Sciences, The Royal Veterinary CollegeClinical Sciences and Services, The Royal Veterinary CollegeComparative Biomedical Sciences, The Royal Veterinary CollegeComparative Biomedical Sciences, The Royal Veterinary CollegeAbstract Background Chronic kidney disease (CKD) is common in geriatric cats, and the most prevalent pathology is chronic tubulointerstitial inflammation and fibrosis. The cell type predominantly responsible for the production of extra-cellular matrix in renal fibrosis is the myofibroblast, and fibroblast to myofibroblast differentiation is probably a crucial event. The cytokine TGF-β1 is reportedly the most important regulator of myofibroblastic differentiation in other species. The aim of this study was to isolate and characterise renal fibroblasts from cadaverous kidney tissue of cats with and without CKD, and to investigate the transcriptional response to TGF-β1. Results Cortical fibroblast cultures were successfully established from the kidney tissue of cats with normal kidney function (FCF) and cats with chronic kidney disease (CKD-FCF). Both cell types expressed the mesenchymal markers vimentin, CD44 and CD29, and were negative for the epithelial marker cytokeratin, mesangial cell marker desmin and endothelial cell marker vWF. Only CKD-FCF expressed VCAM-1, a cell marker associated with inflammation. Incubation with TGF-β1 (0–10 ng/ml) induced a concentration dependent change in cell morphology, and upregulation of myofibroblast marker gene α-SMA expression alongside collagen 1α1, fibronectin, TGF-β1 and CTGF mRNA. These changes were blocked by the TGF-β1 receptor 1 antagonist SB431542 (5 μM). Conclusions FCF and CKD-FCF can be cultured via a simple method and represent a model for the investigation of the progression of fibrosis in feline CKD. The findings of this study suggest TGF-β1 may be involved in fibroblast-myofibroblast transition in feline CKD, as in other species.http://link.springer.com/article/10.1186/s12917-018-1387-2CKDCatsRenal fibrosisMyofibroblast |
| spellingShingle | J. S. Lawson H. M. Syme C. P. D. Wheeler-Jones J. Elliott Characterisation of feline renal cortical fibroblast cultures and their transcriptional response to transforming growth factor β1 BMC Veterinary Research CKD Cats Renal fibrosis Myofibroblast |
| title | Characterisation of feline renal cortical fibroblast cultures and their transcriptional response to transforming growth factor β1 |
| title_full | Characterisation of feline renal cortical fibroblast cultures and their transcriptional response to transforming growth factor β1 |
| title_fullStr | Characterisation of feline renal cortical fibroblast cultures and their transcriptional response to transforming growth factor β1 |
| title_full_unstemmed | Characterisation of feline renal cortical fibroblast cultures and their transcriptional response to transforming growth factor β1 |
| title_short | Characterisation of feline renal cortical fibroblast cultures and their transcriptional response to transforming growth factor β1 |
| title_sort | characterisation of feline renal cortical fibroblast cultures and their transcriptional response to transforming growth factor β1 |
| topic | CKD Cats Renal fibrosis Myofibroblast |
| url | http://link.springer.com/article/10.1186/s12917-018-1387-2 |
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