Volume 4 Supplement 1
Urine-derived Renal Epithelial Cells (URECs) as a source of biomaterial from ciliopathy patients for functional studies and diagnostics
© Ajzeberg et al. 2015
Published: 13 July 2015
Many ciliopathies are diagnosed in infants and children. Obtaining blood and/or skin biopsies for diagnostic and research purposes requires a visit to the clinic and a specialized sample collection skill. We sought alternative non-invasive sources of biomaterial from patients.
We have generated an approved Standard Operating Procedure to isolate and culture primary renal collecting duct cells from urine (URECs).
We show that URECs are approximately 30% proximal tubule cells (megalin positive) and 70% collecting duct cells (AQP2 positive) from the patient. The cells grow on coverslips for immunocytochemistry, can be frozen and grow for 15-20 passages without immortalization. We demonstrate that whereas URECs from healthy individuals ciliate well, URECs from ciliopathy patients with nephronophthisis do not. URECs can form 3D spheroids to study patient-specific tubulogenesis defects. siRNA and overexpression "rescue" experiments in URECs make these cells amenable to functional studies complementing diagnostics. As a proof of principle we show how URECs can be used to test the relevance of genetic variants of "uncertain significance" obtained from whole-exome sequencing. Lastly, we have used patient URECs to explore pharmacological intervention on a patient-specific basis.
This protocol expands the toolkit available to clinical geneticists and researchers alike in a child-friendly manner. Urine culture offers a non-invasive option for genetic and functional testing and does not require the family to go to the clinic for sample donation. We contend that UREC culture will facilitate personalized medicine for the ciliopathy community and beyond.
This article is published under license to BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.