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3D‐electron microscopic characterization of interstitial cells in the human bladder upper lamina propria
Authors:Jochen Neuhaus  Birgit Schröppel  Martin Dass  Hans Zimmermann  Hartwig Wolburg  Petra Fallier‐Becker  Thomas Gevaert  Claus J. Burkhardt  Hoang Minh Do  Jens‐Uwe Stolzenburg
Affiliation:1. Department of Urology, Research Laboratory, University Leipzig, Leipzig, Germany;2. Natural and Medical Sciences Institute (NMI) at the University of Tuebingen, Reutlingen, Germany;3. Carl Zeiss Microscopy GmbH, Training, Application and Support Center (TASC) Application Support EM, Munich, Germany;4. Institute of Pathology and Neuropathology, University Hospital Tuebingen, Tuebingen, Germany;5. Department of Development and Regeneration, KU Leuven, Laboratory of Experimental Urology, Leuven, Belgium;6. Department of Urology, University Leipzig, University Hospital Leipzig, Leipzig, Germany
Abstract:

Aims

To explore the ultrastructure of interstitial cells in the upper lamina propria of the human bladder, to describe the spatial relationships and to investigate cell‐cell contacts.

Methods

Focused ion beam scanning electron microscopy (FIB‐SEM), 3‐View SEM and confocal laser scanning microscopy were used to analyze the 3D ultrastructure of the upper lamina propria in male and female human bladders.

Results

3View‐SEM image stacks as large as 59 × 59 × 17 μm3 (xyz) at a resolution of 16 × 16 × 50 nm3 and high resolution (5 × 5 × 10 nm3) FIB‐SEM stacks could be analyzed. Interstitial cells with myoid differentiation (mIC) and fibroblast like interstitial cells (fIC) were the major cell types in the upper lamina propria. The flat, sheet‐like ICs were oriented strictly parallel to the urothelium. No spindle shaped cells were present. We furthermore identified one branched cell (bIC) with several processes contacting urothelial cells by penetrating the basal membrane. This cell did not make any contacts to other ICs within the upper lamina propria. We found no evidence for the occurrence of telocytes in the upper lamina propria.

Conclusions

Comprehensive 3D‐ultrastructural analysis of the human bladder confirmed distinct subtypes of interstitial cells. We provide evidence for a foremost unknown direct connection between a branched interstitial cell and urothelial cells of which the functional role has still to be elucidated. 3D‐ultrastructure analyses at high resolution are needed to further define the subpopulations of lamina propria cells and cell‐cell interactions.
Keywords:3D‐confocal fluorescence imaging  3D‐scanning electron microscopy (SEM)  cellular fibronectin EDA domain  connexin 43 (Cx43)  Interstitial Cells of Cajal (ICC)  telocyte  wheat germ agglutinin (WGA) staining
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