TY - JOUR
T1 - Characterization of cultured bladder smooth muscle cells
T2 - Assessment of in vitro contractility
AU - Kropp, Bradley P.
AU - Zhang, Yuanyuan
AU - Tomasek, James J.
AU - Cowan, Rick
AU - Furness, Peter D.
AU - Vaughan, Melville B.
AU - Parizi, Mojgan
AU - Cheng, Earl Y.
PY - 1999
Y1 - 1999
N2 - Purpose: The contractile properties of in vitro cultured bladder smooth muscle cells (SMC) are unknown. This study characterized the in vitro contractile response of human and rat bladder SMC to several pharmacological agonists known to induce in vivo contraction of intact bladder muscle. Materials and Methods: Human and rat bladder SMC were seeded separately within attached collagen lattices. Contractility of SMC was analyzed by measuring alterations in lattice diameter after exposure and release to the following contractile agonists: carbachol (10-7 - 10-3 M), calcium- ionophore (10 μM), lysophosphatidic acid (LPA) (1 μM), endothelin (0.1 μM), KCl (3.33 mM) angiotensin II (10 μM), and serotonin (100 μM). Results were recorded as a mean reduction of the lattice diameter. In addition, immunohistochemical analysis for phenotypic markers of smooth muscle cell differentiation was performed on bladder SMC cultured within collagen lattices. Human palmar fascia fibroblasts, which have been previously well characterized by in vitro contractility and immunohistochemistry, were tested in parallel and used as controls for all the above experiments. Results: Human SMC had significant contractile responses to calcium-ionophore (31% ± 4 relative percent contraction, p <0.05), LPA (34% ± 4, p <0.05), and endothelin (37 ± 5%, p <05). There was no significant contraction in response to carbachol, angiotensin II, KCl, or serotonin. Rat bladder SMC had a similar contractile response but did not contract in response to endothelin. In contrast to human and rat bladder SMC, fibroblasts did not contract to calcium-ionophore. Conclusions: In vitro cultured bladder SMC demonstrate loss of contractile response to normal in vivo pharmacologic agonists. Both human and rat bladder SMC can be distinguished in vitro from fibroblasts based upon their lack of contractile response to calcium- ionophore. These results demonstrate the ability to further characterize cultured bladder SMC with in vitro contractility. Further characterization is essential if we are to advance our understanding of the clinical applicability of in vitro studies utilizing cultured bladder SMC.
AB - Purpose: The contractile properties of in vitro cultured bladder smooth muscle cells (SMC) are unknown. This study characterized the in vitro contractile response of human and rat bladder SMC to several pharmacological agonists known to induce in vivo contraction of intact bladder muscle. Materials and Methods: Human and rat bladder SMC were seeded separately within attached collagen lattices. Contractility of SMC was analyzed by measuring alterations in lattice diameter after exposure and release to the following contractile agonists: carbachol (10-7 - 10-3 M), calcium- ionophore (10 μM), lysophosphatidic acid (LPA) (1 μM), endothelin (0.1 μM), KCl (3.33 mM) angiotensin II (10 μM), and serotonin (100 μM). Results were recorded as a mean reduction of the lattice diameter. In addition, immunohistochemical analysis for phenotypic markers of smooth muscle cell differentiation was performed on bladder SMC cultured within collagen lattices. Human palmar fascia fibroblasts, which have been previously well characterized by in vitro contractility and immunohistochemistry, were tested in parallel and used as controls for all the above experiments. Results: Human SMC had significant contractile responses to calcium-ionophore (31% ± 4 relative percent contraction, p <0.05), LPA (34% ± 4, p <0.05), and endothelin (37 ± 5%, p <05). There was no significant contraction in response to carbachol, angiotensin II, KCl, or serotonin. Rat bladder SMC had a similar contractile response but did not contract in response to endothelin. In contrast to human and rat bladder SMC, fibroblasts did not contract to calcium-ionophore. Conclusions: In vitro cultured bladder SMC demonstrate loss of contractile response to normal in vivo pharmacologic agonists. Both human and rat bladder SMC can be distinguished in vitro from fibroblasts based upon their lack of contractile response to calcium- ionophore. These results demonstrate the ability to further characterize cultured bladder SMC with in vitro contractility. Further characterization is essential if we are to advance our understanding of the clinical applicability of in vitro studies utilizing cultured bladder SMC.
KW - Cultured smooth muscle cell
KW - Gel contraction
KW - Human
KW - Rat
KW - Urinary bladder
UR - http://www.scopus.com/inward/record.url?scp=0032881974&partnerID=8YFLogxK
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U2 - 10.1016/S0022-5347(05)68237-7
DO - 10.1016/S0022-5347(05)68237-7
M3 - Article
C2 - 10524934
AN - SCOPUS:0032881974
SN - 0022-5347
VL - 162
SP - 1779
EP - 1784
JO - Journal of Urology
JF - Journal of Urology
IS - 5
ER -