title: Explanting is an ex vivo model of renal epithelial-mesenchymal transition.
abstract: Recognised by their de novo expression of alpha-smooth muscle actin (SMA), recruitment of myofibroblasts is key to the pathogenesis of fibrosis in chronic kidney disease. Increasingly, we realise that epithelial-mesenchymal transition (EMT) may be an important source of these cells. In this study we describe a novel model of renal EMT. Rat kidney explants were finely diced on gelatin-coated Petri dishes and cultured in serum-supplemented media. Morphology and immunocytochemistry were used to identify mesenchymal (vimentin+, α-smooth muscle actin (SMA)+, desmin+), epithelial (cytokeratin+), and endothelial (RECA+) cells at various time points. Cell outgrowths were all epithelial in origin (cytokeratin+) at day 3. By day 10, 50 ± 12% (mean ± SE) of cytokeratin+ cells double-labelled for SMA, indicating EMT. Lectin staining established a proximal tubule origin. By day 17, cultures consisted only of myofibroblasts (SMA+/cytokeratin-). Explanting is a reproducible ex vivo model of EMT. The ability to modify this change in phenotype provides a useful tool to study the regulation and mechanisms of renal tubulointerstitial fibrosis.
authors.affiliation: Department of Nephrology, The Royal Melbourne Hospital, Melbourne, VIC 3050, Australia.
authors.forename: Catherine E
authors.initials: CE
authors.lastname: Winbanks
authors.forename: Ian A
authors.initials: IA
authors.lastname: Darby
authors.forename: Kristen J
authors.initials: KJ
authors.lastname: Kelynack
authors.forename: Dodie
authors.initials: D
authors.lastname: Pouniotis
authors.forename: Gavin J
authors.initials: GJ
authors.lastname: Becker
authors.forename: Tim D
authors.initials: TD
authors.lastname: Hewitson
mesh_terms: D000818:Animals; D015415:Biomarkers / Q000032:analysis; D018929:Cell Culture Techniques; D002454:Cell Differentiation / Q000502:physiology; D049109:Cell Proliferation; D042783:Endothelial Cells / Q000166:cytology / Q000378:metabolism; D004847:Epithelial Cells / Q000166:cytology* / Q000378:metabolism; D058750:Epithelial-Mesenchymal Transition / Q000502:physiology*; D007120:Immunochemistry / Q000379:methods; D007668:Kidney / Q000166:cytology*; D007687:Kidney Tubules, Proximal / Q000378:metabolism*; D059630:Mesenchymal Stem Cells / Q000166:cytology* / Q000378:metabolism; D058628:Myofibroblasts / Q000166:cytology* / Q000378:metabolism; D010641:Phenotype; D051381:Rats; D017207:Rats, Sprague-Dawley; D013194:Staining and Labeling / Q000379:methods
chemical_list: D015415:Biomarkers