Cell News 01/2017
38
Epithelial Morphogenesis of Urine-Derived Renal Epithelial
Cells from Children with Autosomal Recessive Polycystic
Kidney Disease: an Ex Vivo Study
Wolfgang H. Ziegler, Birga Soetje, Arne Mertens, Kathrin Swolana, Margarita E.Georgiadis,
Dieter Haffner
Presenting author: Wolfgang H. Ziegler
Dept. of Paediatric Kidney, Liver and Metabolic Diseases,
Hannover Medical School, Hannover, Germany
Background: Autosomal recessive polycystic kidney disease
(ARPKD) is caused by mutation of the Pkhd1 gene, which
encodes fibrocystin (FPC), a type I membrane protein of largely
unknown function. Among other functions, FPC affects adhe-
sion signaling of cells and their ability to orientate correctly
towards one another. Recently, we established a link between
loss of FPC function and defective epithelial morphogenesis
in 3D cell culture using a canine cell line. Data in humans are
lacking. Therefore, we set up assays for analyzing epithelial
cells from ARPKD patients and healthy controls.
Methods: We take urine-derived renal epithelial cells (URECs)
of ARPKD patients and respective controls in culture. The pa-
tients must have their original kidney(s) and still secrete urine.
Populations of primary cells obtained within 14 days of culture
are being characterized by different criteria and also tested in
3D cell culture conditions, which induce formation of epithelial
spheroids with defined polarity, lumen and cilia.
Results: Using 2D and 3D-culture conditions, we observe
URECs of three different phenotypes. While all different cell
morphologies stain positively for the collecting duct marker
aquaporin 2, only cobblestone-like cells give rise to a strong
epithelial barrier as verified by impedance measurements and
form spheroids in 3D culture conditions. Thus, cells of this
phenotype are being studied preferentially to establish quan-
titative cell parameters with respect to growth rates, transep-
ithelial electric resistance and spheroid formation. Cells of the
suitable phenotype can be collected from all ARPKD patients,
whereas it is restricted in samples from controls requiring a
larger control cohort.
Conclusions: By establishing methods allowing us to charac-
terize the URECs of ARPKD patients, we aim to provide specific
read-outs for future ex vivo analysis of the illness and for
testing options of personalized pharmaceutical intervention.
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