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Abstract: TH-PO559

Spatial Mapping of Cell Populations in the Human Kidney Papilla Using 3D Tissue Cytometry

Session Information

Category: Bone and Mineral Metabolism

  • 401 Bone and Mineral Metabolism: Basic

Authors

  • Makki, Mohammad Shahidul, Indiana University School of Medicine, Indianapolis, Indiana, United States
  • Winfree, Seth, Indiana University School of Medicine, Indianapolis, Indiana, United States
  • Williams, James, Indiana University School of Medicine, Indianapolis, Indiana, United States
  • El-Achkar, Tarek M., Indiana University, Indianapolis, Indiana, United States
Background

Kidney stone disease or nephrolithiasis affects 10% of US population with estimated healthcare cost of more than $10 billion. To date, no clear mechanisms have been identified for the initiation and progression of kidney stone disease. A contributing factor to this knowledge gap is a poor understanding of the various cell types comprising the nephronal segments and the surrounding interstitium in the papilla. To fill this gap, we developed a novel methodology to classify cell types of the papilla based on spatial and morphological features, lectin binding properties which reflects carbohydrate content and staining with Aquaporin 1 (AQP1, detects thin descending limbs and descending vasa recta). The long-term goal is to link subpopulations of cells with other assays such as transcriptome and proteome profiling, and advance our understanding of the renal papillary micro-environment.

Methods

Papillary biopsies were obtained at the time of percutaneous nephrolithotomy or from reference nephrectomy tissues. Fifty-micron thick sections of papilla were stained with fluorescently labelled DBA (Dolichos Biflorus), PNA (Peanut Agglutinin), and/or AQP1. Large scale 3D imaging was performed with confocal microscopy, followed by 3D tissue cytometry analysis using our software tool Volumetric Tissue Exploration and Analysis (VTEA).

Results

With this strategy, VTEA analysis deconvolved all the cells in the papillae into specific subpopulations, which were directly visualized within the tissue. Various subtypes of AQP1+ thin descending limb cells or AQP1- thin ascending limb cells were classified based on the presence of co-labeling with DBA and/or PNA. Collecting ducts and ducts of Bellini, which are easily identified by morphological features, stained predominantly with PNA. AQP1 weak staining cells that did not stain with any lectins were identified as vascular cells of the vasa recta.

Conclusion

Using a straightforward fluorescence based staining strategy, we can distinguish subpopulations of epithelial and vascular cells in the papilla. Implementing this strategy to link cell subtypes with “omics” signatures will define the biological significance of these subpopulation, and advance our understanding of the complex homeostasis within the papilla and its perturbance by stone disease

Funding

  • NIDDK Support