Journal article
Porcine bladder urothelial, myofibroblast, and detrusor muscle cells: Characterization and ATP release
Frontiers in Pharmacology, Vol.2, 27
2011
Abstract
ATP is released from the bladder mucosa in response to stretch, but the cell types responsible are unclear. Our aim was to isolate and characterize individual populations of urothelial, myofibroblast, and detrusor muscle cells in culture, and to examine agonist-stimulated ATP release. Using female pig bladders, urothelial cells were isolated from bladder mucosa following trypsin-digestion of the luminal surface. The underlying myofibroblast layer was dissected, minced, digested, and cultured until confluent (10-14 days). A similar protocol was used for muscle cells. Cultures were used for immunocytochemical staining and/or ATP release investigations. In urothelial cultures, immunoreactivity was present for the cytokeratin marker AE1/AE3 but not the contractile protein α-smooth muscle actin (α-SMA) or the cytoskeletal filament vimentin. Neither myofibroblast nor muscle cell cultures stained for AE1/AE3. Myofibroblast cultures partially stained for α-SMA, whereas muscle cultures were 100% stained. Both myofibroblast and muscle stained for vimentin, however, they were morphologically distinct. Ultrastructural studies verified that the suburothelial layer of pig bladder contained abundant myofibroblasts, characterized by high densities of rough endoplasmic reticulum. Baseline ATP release was higher in urothelial and myofibroblast cultures, compared with muscle. ATP release was significantly stimulated by stretch in all three cell populations. Only urothelial cells released ATP in response to acid, and only muscle cells were stimulated by capsaicin. Tachykinins had no effect on ATP release. In conclusion, we have established a method for culture of three cell populations from porcine bladder, a well-known human bladder model, and shown that these are distinct morphologically, immunologically, and pharmacologically. © 2011 Cheng, Mansfield, Sandow, Sadananda, Burcher and Moore.
Details
- Title
- Porcine bladder urothelial, myofibroblast, and detrusor muscle cells: Characterization and ATP release
- Authors
- Y Cheng (Author) - University of New South WalesK J Mansfield (Author) - University of WollongongShaun L Sandow (Author) - University of New South WalesP Sadananda (Author) - University of New South WalesE Burcher (Author) - University of New South WalesK H Moore (Author) - University of New South Wales
- Publication details
- Frontiers in Pharmacology, Vol.2, 27; 9
- Publisher
- Frontiers Research Foundation
- Date published
- 2011
- DOI
- 10.3389/fphar.2011.00027
- ISSN
- 1663-9812
- Copyright note
- Copyright © 2011 The Authors. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
- Organisation Unit
- School of Health - Biomedicine; University of the Sunshine Coast, Queensland; School of Health and Sport Sciences - Legacy; School of Health and Behavioural Sciences - Legacy
- Language
- English
- Record Identifier
- 99449057702621
- Output Type
- Journal article
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