Immortalized human bladder cell line exhibits amiloride-sensitive sodium absorption

Perrone, Ron; Johns, Conrado; Grubman, Shelley A.; Moy, E.; Lee, D. W.; Alroy, Joseph; Sant, Grannum R.; Jefferson, Douglas M.

doi:10.1152/ajprenal.1996.270.1.f148

Cited by 6 publications

(7 citation statements)

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“…Perrone and colleagues (33) also assessed the electrophysiological properties of human urothelial cells in culture, but they achieved a mean TER of only 576 ⍀⅐cm 2 , an order of magnitude below that recorded in the present study. Whereas NHU cell lines in this study were obtained from patients with no history or evidence of urothelial pathology, Perrone and Fig.…”

Section: Since the First Serial Cultivation Of Nhu Cells In Vitrocontrasting

confidence: 74%

“…Nevertheless, it has been proposed that increased urothelial permeability has a role in the pathophysiology of interstitial cystitis (32). Thus the difference in TER values obtained in this and Perrone's study (33) could be due to an inherent dysfunction in the urothelium in interstitial cystitis and indicates the potential value of in vitro models for studying interstitial cystitis and other such associated conditions.…”

Section: Since the First Serial Cultivation Of Nhu Cells In Vitromentioning

confidence: 51%

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A biomimetic tissue from cultured normal human urothelial cells: analysis of physiological function

Cross

Eardley²,

Leese³

et al. 2005

American Journal of Physiology-Renal Physiology

109

126

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The urinary bladder and associated tract is lined by the urothelium. Once considered as just an impermeable epithelium, it is becoming evident that the urothelium not only functions as a volume-accommodating urinary barrier but has additional roles, including sensory signaling. Lack of access to normal human urothelium has hampered physiological investigation, and although cell culture systems have been developed, there has been a failure to demonstrate that normal human urothelial (NHU) cells grown in vitro retain the capacity to form a functional differentiated urothelium. The aim of this study was to develop a biomimetic human urothelium from NHU cell cultures. Urothelial cells isolated from normal human urothelium and serially propagated as monolayers in serum-free culture were homogeneous and adopted a proliferative, nondifferentiated phenotype. In the presence of serum and physiological concentrations of calcium, these cells could be reproducibly induced to form stratified urothelia consisting of basal, intermediate, and superficial cells, with differential expression of cytokeratins and superficial tight junctions. Functionally, the neotissues showed characteristics of native urothelium, including high transepithelial electrical resistance of >3,000 Omega.cm(2), apical membrane-restricted amiloride-sensitive sodium ion channels, basal expression of Na(+)-K(+)-ATPase, and low diffusive permeability to urea, water, and dextran. This model represents major progress in developing a biomimetic human urothelial culture model to explore molecular and functional relationships in normal and dysfunctional bladder physiology.

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Section: Since the First Serial Cultivation Of Nhu Cells In Vitrocontrasting

confidence: 74%

Section: Since the First Serial Cultivation Of Nhu Cells In Vitromentioning

confidence: 51%

A biomimetic tissue from cultured normal human urothelial cells: analysis of physiological function

Cross

Eardley²,

Leese³

et al. 2005

American Journal of Physiology-Renal Physiology

109

126

View full text Add to dashboard Cite

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“…However, they presented no evidence on the expression of differentiation markers or the achievement of barrier function (23). Finally, Perrone et al (37) have recently described the establishment of an immortalized human bladder cell line from a patient with interstitial cystitis. This multilayer culture model was the only system described to date that formed reasonably tight monolayers (TER of 500 -1000 ⍀ cm 2 ) and contained amiloride-sensitive apical sodium absorption.…”

Section: Discussionmentioning

confidence: 99%

Primary Uroepithelial Cultures

Truschel¹,

Ruiz²,

Shul'man³

et al. 1999

Journal of Biological Chemistry

102

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Despite almost 25 years of effort, the development of a highly differentiated and functionally equivalent cell culture model of uroepithelial cells has eluded investigators. We have developed a primary cell culture model of rabbit uroepithelium that consists of an underlying cell layer that interacts with a collagen substratum, an intermediate cell layer, and an upper cell layer of large (25-100 m) superficial cells. When examined at the ultrastructural level, the superficial cells formed junctional complexes and had an asymmetric unit membrane, a hallmark of terminal differentiation in bladder umbrella cells. These cultured "umbrella" cells expressed uroplakins and a 27-kDa uroepithelial specific antigen that assembled into detergent-resistant asymmetric unit membrane particles. The cultures had low diffusive permeabilities for water (2.8 ؋ 10 ؊4 cm/s) and urea (3.0 ؋ 10 ؊7 cm/s) and high transepithelial resistance (>8000 ⍀ cm 2 ) was achieved when 1 mM CaCl 2 was included in the culture medium. The cell cultures expressed an amiloride-sensitive sodium transport pathway and increases in apical membrane capacitance were observed when the cultures were osmotically stretched. The described primary rabbit cell culture model mimics many of the characteristics of uroepithelium found in vivo and should serve as a useful tool to explore normal uroepithelial function as well as dysfunction as a result of disease.

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“…In only two studies of urothelial tissue culture were some or most of the above criteria filled. Perrone and colleagues (37) immortalized and grew human urothelial cells on permeable supports. These tissue culture cells had a transepithelial resistance of 500-1,000 ⍀ •cm 2 , an amiloride-sensitive current of 2 µA/cm 2 , and were four cell layers thick and characterized by the presence of tight junctions.…”

Section: Inertnessmentioning

confidence: 99%

Everything you wanted to know about the bladder epithelium but were afraid to ask

Lewis

2000

American Journal of Physiology-Renal Physiology

298

247

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The mammalian urinary bladder epithelium (urothelium) performs the important function of storing urine for extended periods, while maintaining the urine composition similar to that delivered by the kidneys. The urothelium possesses four properties to perform this function. First, it offers a minimum epithelial surface area-to-urine volume; this reduces the surface area for passive movement of substances between lumen and blood. Second, the passive permeability of the apical membrane and tight junctions is very low to electrolytes and nonelectrolytes. Third, the urothelium has a hormonally regulated sodium absorptive system; thus passive movement of sodium from blood to urine is countered by active sodium reabsorption. Last, the permeability properties of the apical membrane and tight junctions of the urothelium are not altered by most substances found in the urine or blood. The importance of the barrier function of the urothelium is illustrated by infectious cystitis. The loss of the barrier function results in the movement of urinary constituents into the lamina propria and underlying muscle layers, resulting in suprapubic and lower back pain and frequent, urgent, and painful voiding.

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Immortalized human bladder cell line exhibits amiloride-sensitive sodium absorption

Cited by 6 publications

References 0 publications

A biomimetic tissue from cultured normal human urothelial cells: analysis of physiological function

A biomimetic tissue from cultured normal human urothelial cells: analysis of physiological function

Primary Uroepithelial Cultures

Everything you wanted to know about the bladder epithelium but were afraid to ask

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