Primary Cultures of Human Urothelial Cells for Genotoxicity Testing

Flieger, A.; Golka, Klaus; Schulze, Harald; Föllmann, Wolfram

doi:10.1080/15287390801988939

Cited by 17 publications

(15 citation statements)

References 18 publications

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“…First, the use of patient-derived cells requires secondary surgical procedures for tissue isolation and propagation of in vitro cultures [5], thereby requiring substantial laboratory resources to attain sufficient cell numbers for implant seeding. In addition, urothelial and smooth muscle cells are subject to de-differentiation during in vitro cultivation [11,12], potentially compromising the in vivo performance of seeded constructs. Moreover, autologous cells isolated from diseased bladders have been shown to display various functional abnormalities [13,14] and therefore may not be suitable for restoration of normal organ operations.…”

Section: Introductionmentioning

confidence: 99%

Bladder tissue regeneration using acellular bi-layer silk scaffolds in a large animal model of augmentation cystoplasty

Chung

Gil

et al. 2013

Biomaterials

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A cellular scaffolds derived from Bombyx mori silk fibroin were investigated for their ability to support functional tissue regeneration in a porcine model of augmentation cystoplasty. Two bi-layer matrix configurations were fabricated by solvent-casting/salt leaching either alone (Group 1) or in combination with silk film casting (Group 2) to yield porous foams buttressed by heterogeneous surface pore occlusions or homogenous silk films, respectively. Bladder augmentation was performed with each scaffold group (6×6cm2) in juvenile Yorkshire swine for 3 m of implantation. Augmented animals exhibited high rates of survival (Group 1: 5/6, 83%; Group 2: 4/4, 100%) and voluntary voiding over the course of the study period. Urodynamic evaluations demonstrated mean increases in bladder capacity over pre-operative levels (Group 1: 277%; Group 2: 153%) which exceeded non surgical control gains (144%) encountered due to animal growth. Similarly, elevations in bladder compliance were substantially higher in augmented animals from baseline (Group 1: 357%; Group 2: 147%) in comparison to controls (41%). Gross tissue evaluations revealed that both matrix configurations supported extensive de novo tissue formation throughout the entire original implantation site which exhibited ultimate tensile strength similar to nonsurgical counterparts. Histological and immunohistochemical analyses showed that both implant groups promoted comparable extents of smooth muscle regeneration and contractile protein (α-smooth muscle actin and SM22α) expression within defect sites similar to controls. Parallel evaluations demonstrated the formation of a transitional, multi-layered urothelium with prominent cytokeratin, uroplakin, and p63 protein expression in both matrix groups. De novo innervation and vascularization processes were evident in all regenerated tissues indicated by synaptophysin-positive neuronal cells and vessels lined with CD31 expressing endothelial cells. Ex vivo organ bath studies demonstrated that regenerated tissues supported by both silk matrices displayed contractile responses to carbachol, α,β-methylene-ATP, KCl, and electrical field stimulation similar to controls. Our data detail the ability of acellular silk scaffolds to support regeneration of innervated, vascularized smooth muscle and urothelial tissues within 3 m with structural, mechanical, and functional properties comparable to native tissue in a porcine model of bladder repair.

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Section: Introductionmentioning

confidence: 99%

Bladder tissue regeneration using acellular bi-layer silk scaffolds in a large animal model of augmentation cystoplasty

Chung

Gil

et al. 2013

Biomaterials

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“…The immunostaining results demonstrated that on PLCL and PS, CK7/8 and CK19 staining was evident during the whole cell culture period as expected, because those markers are present in all layers of normal urothelium [14,[22][23][24]. In contrast, on the hAM, the CK19 expression of hUCs decreased with time.…”

Section: Discussionmentioning

confidence: 92%

Comparison of a poly- l -lactide-co- ɛ -caprolactone and human amniotic membrane for urothelium tissue engineering applications

Sartoneva

Haimi

Miettinen

et al. 2010

J. R. Soc. Interface.

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The reconstructive surgery of urothelial defects, such as severe hypospadias is susceptible to complications. The major problem is the lack of suitable grafting materials. Therefore, finding alternative treatments such as reconstruction of urethra using tissue engineering is essential. The aim of this study was to compare the effects of naturally derived acellular human amniotic membrane (hAM) to synthetic poly-L-lactide-co-1-caprolactone (PLCL) on human urothelial cell (hUC) viability, proliferation and urothelial differentiation level. The viability of cells was evaluated using live/dead staining and the proliferation was studied using WST-1 measurement. Cytokeratin (CK)7/8 and CK19 were used to confirm that the hUCs maintained their phenotype on different biomaterials. On the PLCL, the cell number significantly increased during the culturing period, in contrast to the hAM, where hUC proliferation was the weakest at 7 and 14 days. In addition, the majority of cells were viable and maintained their phenotype when cultured on PLCL and cell culture plastic, whereas on the hAM, the viability of hUCs decreased with time and the cells did not maintain their phenotype. The PLCL membranes supported the hUC proliferation significantly more than the hAM. These results revealed the significant potential of PLCL membranes in urothelial tissue engineering applications.

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“…Samples JSCRT Volume-2 | Issue-1 August, 2018 collected from younger patients had better urothelial cell growth in culture than samples collected from older patients. Other studies have demonstrated that age factor influenced the rate of urothelial cell growth [4,5,6] .…”

Section: Discussionmentioning

confidence: 99%

“…A typical human urothelial cell culture model system could help the experimental investigation of urothelial wound repair. Studies involving scratch wounding of confluent urothelial cell monolayers have been useful in identifying critical signaling pathways [4][5][6][7] . Studies on the usage of adult stem cells to treat patients with severe burns and hematological diseases have been successful [5,6] .…”

Section: Introductionmentioning

confidence: 99%

Human Urothelial Cells Isolation, In Vitro Expansion and Characterization for Evaluating Bio-Engineering Potentials

Lamplot

Qin²,

Guo³

et al. 2018

JSCRT

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Urothelial damage results in compromised urothelial barrier function and leakage of urine causing frequent bladder infections. The urothelium exhibits one of the lowest turnover rates among mammalian epithelia under homeostatic conditions. To explore the bio-engineering possibility to restore the urothelial lining of the urinary tract, we aimed to establish in vitro culture and characterization of urothelial cells from human ureter biopsies. In vitro cultures of urothelial cells were characterized for CK7, CK20, Ki67, Ecadherin, AE1, Uroplakin III marker expression. When in vitro culture of urothelial cells was maintained in low calcium (0.09mM) conditions, cells in culture expressed CK7, AE1, Ki67, low expression of E-cadherin and negative for uroplakins III, CK20 markers. However, when urothelial cells were exposed to 2mM calcium concentration, in vitro grown urothelial cells showed positive expression for CK7, AE1, Uroplakin III, E-cadherin and mild expression of CK20, Ki67 markers. Cultured urothelial cells can respond to in vitro differentiation cues. These in vitro expanded urothelial cell cultures, optimized in serum-free cell culture media and systematically characterized urothelial cells may be a prelude to in vivo bio-engineering applications in planned mice models.

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Primary Cultures of Human Urothelial Cells for Genotoxicity Testing

Cited by 17 publications

References 18 publications

Bladder tissue regeneration using acellular bi-layer silk scaffolds in a large animal model of augmentation cystoplasty

Bladder tissue regeneration using acellular bi-layer silk scaffolds in a large animal model of augmentation cystoplasty

Comparison of a poly- l -lactide-co- ɛ -caprolactone and human amniotic membrane for urothelium tissue engineering applications

Human Urothelial Cells Isolation, In Vitro Expansion and Characterization for Evaluating Bio-Engineering Potentials

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