Characterization of the control of intracellular [Ca2+] and the contractile phenotype of cultured human detrusor smooth muscle cells

The Journal of Urology
D N WoodC H Fry

Abstract

We measured the functional properties of cultured human detrusor myocytes with respect to their ability to regulate their intracellular [Ca2+] and generate force in collagen matrices. Human detrusor biopsies were dissociated into single cells by collagenase treatment and used immediately or cultured in D-valine medium and subsequently used after culture trypsinization. Intracellular [Ca2+] was measured in Fura-2 loaded myocytes. Cell force development was measured by incorporating cells into a collagen gel and attaching it to an isometric strain gauge. Carbachol was equally effective in generating Ca transients in freshly isolated and cultured cells. Carbachol potency (pEC50) and the magnitude of Ca2+ transients were similar. Adenosine triphosphate potency was decreased in cultured cells and Ca2+ transients showed properties consistent with a purinoceptor shift from a purinergic subtype. Temporal restitution of Ca2+ transients was similar in the 2 groups, indicative of retained intracellular Ca2+ stores in cultured cells. Cultured cells (approximately 10(6)) embedded in collagen gel generated a force about 10 times greater than that generated by gel alone. The cell dependent force could be further increased by adding carbachol....Continue Reading

References

Dec 1, 1991·The American Journal of Physiology·C M YangH J Chien
Jul 1, 1996·Experimental Physiology·P ChambersJ I Gillespie
Mar 19, 1999·Experimental Physiology·P ChambersJ I Gillespie
Jun 11, 2002·American Journal of Physiology. Cell Physiology·Yongmu ZhengMichael E DiSanto
Apr 5, 2003·American Journal of Physiology. Regulatory, Integrative and Comparative Physiology·N ThiruchelvamC H Fry

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Citations

Sep 8, 2011·BJU International·Felix WezelDavid F M Thomas
Jun 11, 2011·Neurourology and Urodynamics·C H FryR Clayton
Oct 4, 2008·Current Opinion in Urology·Dan Wood, Jennifer Southgate

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