Porcine bladder urothelial, myofibroblast and detrusor muscle cells: characterisation and ATP release
ATP is released from the bladder mucosa in response to stretch, but the cell types responsible are unclear. Our aim was to isolate and characterise individual populations of urothelial, myofibroblast and detrusor muscle cells in culture, and to examine agonist-stimulated ATP release. Using female pi...
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| Format: | Article |
| Language: | English |
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Frontiers Media S.A.
2011-06-01
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| Series: | Frontiers in Pharmacology |
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| Online Access: | http://journal.frontiersin.org/Journal/10.3389/fphar.2011.00027/full |
| _version_ | 1819142542907670528 |
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| author | Ying eCheng Kylie eMansfield Shaun eSandow Prajni eSadananda Elizabeth eBurcher Kate eMoore |
| author_facet | Ying eCheng Kylie eMansfield Shaun eSandow Prajni eSadananda Elizabeth eBurcher Kate eMoore |
| author_sort | Ying eCheng |
| collection | DOAJ |
| description | ATP is released from the bladder mucosa in response to stretch, but the cell types responsible are unclear. Our aim was to isolate and characterise 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, characterised 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. |
| first_indexed | 2024-12-22T12:12:01Z |
| format | Article |
| id | doaj.art-afe10595e39549958c4ed9223ee6d499 |
| institution | Directory Open Access Journal |
| issn | 1663-9812 |
| language | English |
| last_indexed | 2024-12-22T12:12:01Z |
| publishDate | 2011-06-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Pharmacology |
| spelling | doaj.art-afe10595e39549958c4ed9223ee6d4992022-12-21T18:26:16ZengFrontiers Media S.A.Frontiers in Pharmacology1663-98122011-06-01210.3389/fphar.2011.0002710618Porcine bladder urothelial, myofibroblast and detrusor muscle cells: characterisation and ATP releaseYing eCheng0Kylie eMansfield1Shaun eSandow2Prajni eSadananda3Elizabeth eBurcher4Kate eMoore5University of New South WalesUniversity of WollongongUniversity of New South WalesUniversity of New South WalesUniversity of New South WalesUniversity of New South WalesATP is released from the bladder mucosa in response to stretch, but the cell types responsible are unclear. Our aim was to isolate and characterise 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, characterised 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.http://journal.frontiersin.org/Journal/10.3389/fphar.2011.00027/fullUrotheliumATPBladderdetrusorultrastructuremyofibroblast |
| spellingShingle | Ying eCheng Kylie eMansfield Shaun eSandow Prajni eSadananda Elizabeth eBurcher Kate eMoore Porcine bladder urothelial, myofibroblast and detrusor muscle cells: characterisation and ATP release Frontiers in Pharmacology Urothelium ATP Bladder detrusor ultrastructure myofibroblast |
| title | Porcine bladder urothelial, myofibroblast and detrusor muscle cells: characterisation and ATP release |
| title_full | Porcine bladder urothelial, myofibroblast and detrusor muscle cells: characterisation and ATP release |
| title_fullStr | Porcine bladder urothelial, myofibroblast and detrusor muscle cells: characterisation and ATP release |
| title_full_unstemmed | Porcine bladder urothelial, myofibroblast and detrusor muscle cells: characterisation and ATP release |
| title_short | Porcine bladder urothelial, myofibroblast and detrusor muscle cells: characterisation and ATP release |
| title_sort | porcine bladder urothelial myofibroblast and detrusor muscle cells characterisation and atp release |
| topic | Urothelium ATP Bladder detrusor ultrastructure myofibroblast |
| url | http://journal.frontiersin.org/Journal/10.3389/fphar.2011.00027/full |
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