Abstract
Identification of biochemical and mechanical stimuli in order to modulate the function of bladder smooth muscle cells (SMC) in viable detrusor constructs. Human bladder detrusor cells were seeded on bladder acellular matrix and cultured under different conditions. Cell viability and proliferation were assessed by fluorescent microscopic analyses. Histological, immunohistochemical and flow cytometric analyses were performed to compare growth characteristics and differentiation of SMC. The combination of medium conditioned with proliferative urothelium and mechanical stretch resulted in a more densely populated membrane. In this culture system, the expression of α-smooth muscle actin (α-SMA) and desmin were clearly induced after serum elimination. SMC-phenotype can be modulated in viable detrusor constructs by applying selected combinations of urothelial-conditioned media and mechanical stimulation under stepwise reduction and elimination of serum.
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Abbreviations
- SMC:
-
Bladder smooth muscle cells
- P3:
-
Bladder detrusor cells of passage 3
- FCS:
-
Fetal calf serum
- α-SMA:
-
α-smooth muscle actin
- H&E:
-
Hematoxylin and Eosin staining
- DAPI:
-
4’-6-Diamidino-2-phenylindole
- BCECF:
-
2’,7’-Bis-carboxyethyl-5 and 6-carboxyfluorescein
- PI:
-
Propidium iodide
References
Albinsson S, Nordstrom I, Hellstrand P (2004) Stretch of the vascular wall induces smooth muscle differentiation by promoting actin polymerization. J Biol Chem 279:34849–34855
Bottger BA, Hedin U, Johansson S, Thyberg J (1989) Integrin-type fibronectin receptors of rat arterial smooth muscle cells: isolation, partial characterization and role in cytoskeletal organization and control of differentiated properties. Differentiation 41:158–167
Camoretti-Mercado B, Liu HW, Halayko AJ, Forsythe SM, Kyle JW, Li B (2000) Physiological control of smooth muscle-specific gene expression through regulated nuclear translocation of serum response factor. J Biol Chem 275:30387–30393
Davidson RA, McCloskey KD (2005) Morphology and localization of interstitial cells in the ginea pig bladder: structural relationships with smooth muscle and neurons. J Urol 173:1385–1390
Drake MJ, Hedlund P, Andersson KE, Brading AF, Hussain I, Fowler C (2003) Morphology, phenotype and ultrastructure of fibroblastic cells from normal and neuropathic human detrusor: absence of myofibroblast characteristics. J Urol 169:1573–1576
Fossum M, Nordenskjold A, Kratz G (2004) Engineering of multilayered urinary tissue in vitro. Tissue Eng 10:175–180
Galvin DJ, Watson RW, Gillespie JI, Brady H, Fitzpatrick JM (2002) Mechanical stretch regulates cell survival in human bladder smooth muscle cells in vitro. Am J Physiol Renal Physiol 283:F1192–1199
Hashitani H, Yanai Y, Suzuki H (2004) Role of interstitial cells and gap junctions in the transmission of spontaneous Ca2+ signals in detrusor smooth muscles of the guinea-pig urinary bladder. J Physiol 559:567–581
Hedin U, Bottger BA, Forsberg E, Johansson S, Thyberg J (1988) Diverse effects of fibronectin and laminin on phenotypic properties of cultured arterial smooth muscle cells. J Cell Biol 107:307–319
Hedin U, Bottger BA, Luthman J, Johansson S, Thyberg J (1989) A substrate of the cell-attachment sequence of fibronectin (Arg-Gly-Asp-Ser) is sufficient to promote transition of arterial smooth muscle cells from a contractile to a synthetic phenotype. Dev Biol 133:489–501
Kropp BP, Zhang Y, Tomasek JJ, Cowan R, Furness PD 3rd, Vaughan MB et al (1999) Characterization of cultured bladder smooth muscle cells: assessment of in vitro contractility. J Urol 162:1779–1784
Lai JY, Yoon CY, Yoo JJ, Wulf T, Atala A (2002) Phenotypic and functional characterization of in vivo tissue engineered smooth muscle from normal and pathological bladders. J Urol 168:1853–1857
Liu W, Li Y, Cunha S, Hayward G, Baskin L (2000) Diffusable growth factors induce bladder smooth muscle differentiation. In Vitro Cell Dev Biol Anim 36:476–484
Ram-Liebig G, Meye A, Hakenberg OW, Haase M, Baretton G, Wirth MP (2004) Induction of proliferation and differentiation of cultured urothelial cells on acellular biomaterials. BJU Int 94:922–927
Schultheiss D, Lorenz RR, Meister R, Westphal M, Gabouev AI, Mertsching H, Biancosino C, Schlote N, Wefer J, Winkler M, Stief CG, Jonas U (2004) Functional tissue engineering of autologous tunica albuginea: a possible graft for Peyronie’s disease surgery. Eur Urol 45:781–786
Sjuve R, Haase H, Ekblad E, Malmqvist U, Morano I, Arner A (2001) Increased expression of non-muscle myosin heavy chain-B in connective tissue cells of hypertrophic rat urinary bladder. Cell Tissue Res 304:271–278
Yoo JJ, Meng J, Oberpenning F, Atala A (1998) Bladder augmentation using allogenic bladder submucosa seeded with cells. Urology 51:221–225
Zeidan A, Nordstrom I, Albinsson S, Malmqvist U, Sward K, Hellstrand P (2003) Stretch-induced contractile differentiation of vascular smooth muscle: sensitivity to actin polymerization inhibitors. Am J Physiol Cell Physiol 284:C1387–1396
Zhang Y, Kropp BP, Lin HK, Cowan R, Cheng EY (2004) Bladder regeneration with cell-seeded small intestinal submucosa. Tissue Eng 10:181–187
Zhang Y, Kropp BP, Moore P, Cowan R, Furness PD 3rd, Kolligian ME, Frey P, Cheng EY (2000) Coculture of bladder urothelial and smooth muscle cells on small intestinal submucosa: potential applications for tissue engineering technology. J Urol 164:928–934
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Ram-Liebig, G., Ravens, U., Balana, B. et al. New approaches in the modulation of bladder smooth muscle cells on viable detrusor constructs. World J Urol 24, 429–437 (2006). https://doi.org/10.1007/s00345-006-0104-0
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DOI: https://doi.org/10.1007/s00345-006-0104-0