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Immunocytochemical characterisation of cultures of human bladder mucosal cells.

Woodman JR, Mansfield KJ, Lazzaro VA, Lynch W, Burcher E, Moore KH - BMC Urol (2011)

Bottom Line: Individual populations of urothelial and myofibroblast-like cells were isolated using magnetic activated cell separation (MACS).Cells were slow growing, needing 3 to 5 weeks to attain confluence.This may be important pathologically but also suggests that studies of the physiological function of these cells in culture may not give a reliable indicator of human physiology.

View Article: PubMed Central - HTML - PubMed

Affiliation: Detrusor Muscle Laboratory, The St George Hospital, University of New South Wales, Sydney, NSW 2052, Australia.

ABSTRACT

Background: The functional role of the bladder urothelium has been the focus of much recent research. The bladder mucosa contains two significant cell types: urothelial cells that line the bladder lumen and suburothelial interstitial cells or myofibroblasts. The aims of this study were to culture these cell populations from human bladder biopsies and to perform immunocytochemical characterisation.

Methods: Primary cell cultures were established from human bladder biopsies (n = 10). Individual populations of urothelial and myofibroblast-like cells were isolated using magnetic activated cell separation (MACS). Cells were slow growing, needing 3 to 5 weeks to attain confluence.

Results: Cytokeratin 20 positive cells (umbrella cells) were isolated at primary culture and also from patients' bladder washings but these did not proliferate. In primary culture, proliferating cells demonstrated positive immunocytochemical staining to cytokeratin markers (AE1/AE3 and A0575) as well fibroblasts (5B5) and smooth muscle (αSMA) markers. An unexpected finding was that populations of presumptive urothelial and myofibroblast-like cells, isolated using the MACS beads, stained for similar markers. In contrast, staining for cytokeratins and fibroblast or smooth muscle markers was not co-localised in full thickness bladder sections.

Conclusions: Our results suggest that, in culture, bladder mucosal cells may undergo differentiation into a myoepithelial cell phenotype indicating that urothelial cells have the capacity to respond to environmental changes. This may be important pathologically but also suggests that studies of the physiological function of these cells in culture may not give a reliable indicator of human physiology.

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Bladder superficial urothelial cells isolated from bladder washings. Phase contrast microscopy demonstrated large spreading cells (A) which were positive for the umbrella cell marker CK20 (B). (Bar = 50 μm)
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Figure 3: Bladder superficial urothelial cells isolated from bladder washings. Phase contrast microscopy demonstrated large spreading cells (A) which were positive for the umbrella cell marker CK20 (B). (Bar = 50 μm)

Mentions: The cells isolated from the bladder washings were large spreading cells (Figure 3A) that were positive for A0575 and CK20, a marker indicative of fully differentiated superficial umbrella cells(Figure 3B). At the time of isolation the isolated umbrella cells were able to exclude trypan blue indicating cell viability. However, similar to the CK20-positive cells isolated from bladder biopsy specimens, these cells did not proliferate in culture.


Immunocytochemical characterisation of cultures of human bladder mucosal cells.

Woodman JR, Mansfield KJ, Lazzaro VA, Lynch W, Burcher E, Moore KH - BMC Urol (2011)

Bladder superficial urothelial cells isolated from bladder washings. Phase contrast microscopy demonstrated large spreading cells (A) which were positive for the umbrella cell marker CK20 (B). (Bar = 50 μm)
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3104367&req=5

Figure 3: Bladder superficial urothelial cells isolated from bladder washings. Phase contrast microscopy demonstrated large spreading cells (A) which were positive for the umbrella cell marker CK20 (B). (Bar = 50 μm)
Mentions: The cells isolated from the bladder washings were large spreading cells (Figure 3A) that were positive for A0575 and CK20, a marker indicative of fully differentiated superficial umbrella cells(Figure 3B). At the time of isolation the isolated umbrella cells were able to exclude trypan blue indicating cell viability. However, similar to the CK20-positive cells isolated from bladder biopsy specimens, these cells did not proliferate in culture.

Bottom Line: Individual populations of urothelial and myofibroblast-like cells were isolated using magnetic activated cell separation (MACS).Cells were slow growing, needing 3 to 5 weeks to attain confluence.This may be important pathologically but also suggests that studies of the physiological function of these cells in culture may not give a reliable indicator of human physiology.

View Article: PubMed Central - HTML - PubMed

Affiliation: Detrusor Muscle Laboratory, The St George Hospital, University of New South Wales, Sydney, NSW 2052, Australia.

ABSTRACT

Background: The functional role of the bladder urothelium has been the focus of much recent research. The bladder mucosa contains two significant cell types: urothelial cells that line the bladder lumen and suburothelial interstitial cells or myofibroblasts. The aims of this study were to culture these cell populations from human bladder biopsies and to perform immunocytochemical characterisation.

Methods: Primary cell cultures were established from human bladder biopsies (n = 10). Individual populations of urothelial and myofibroblast-like cells were isolated using magnetic activated cell separation (MACS). Cells were slow growing, needing 3 to 5 weeks to attain confluence.

Results: Cytokeratin 20 positive cells (umbrella cells) were isolated at primary culture and also from patients' bladder washings but these did not proliferate. In primary culture, proliferating cells demonstrated positive immunocytochemical staining to cytokeratin markers (AE1/AE3 and A0575) as well fibroblasts (5B5) and smooth muscle (αSMA) markers. An unexpected finding was that populations of presumptive urothelial and myofibroblast-like cells, isolated using the MACS beads, stained for similar markers. In contrast, staining for cytokeratins and fibroblast or smooth muscle markers was not co-localised in full thickness bladder sections.

Conclusions: Our results suggest that, in culture, bladder mucosal cells may undergo differentiation into a myoepithelial cell phenotype indicating that urothelial cells have the capacity to respond to environmental changes. This may be important pathologically but also suggests that studies of the physiological function of these cells in culture may not give a reliable indicator of human physiology.

Show MeSH
Related in: MedlinePlus