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Human proximal tubule cells form functional microtissues.

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Prange, Jenny A Bieri, Manuela Segerer, Stephan Burger, Charlotte Kaech, Andres Devuyst, Olivier [UCL]

The epithelial cells lining the proximal tubules of the kidney mediate complex transport processes and are particularly vulnerable to drug toxicity. Drug toxicity studies are classically based on two-dimensional cultures of immortalized proximal tubular cells. Such immortalized cells are dedifferentiated, and lose transport properties (including saturable endocytic uptake) encountered in vivo. Generating differentiated, organotypic human microtissues would potentially alleviate these limitations and facilitate drug toxicity studies. Here, we describe the generation and characterization of kidney microtissues from immortalized (HK-2) and primary (HRPTEpiC) human renal proximal tubular epithelial cells under well-defined conditions. Microtissue cultures were done in hanging drop GravityPLUS™ culture plates and were characterized for morphology, proliferation and differentiation markers, and by monitoring the endocytic uptake of albumin. Kidney microtissues were successfully obtained by co-culturing HK-2 or HRPTEpiC cells with fibroblasts. The HK-2 microtissues formed highly proliferative, but dedifferentiated microtissues within 10 days of culture, while co-culture with fibroblasts yielded spherical structures already after 2 days. Low passage HRPTEpiC microtissues (mono- and co-culture) were less proliferative and expressed tissue-specific differentiation markers. Electron microscopy evidenced epithelial differentiation markers including microvilli, tight junctions, endosomes, and lysosomes in the co-cultured HRPTEpiC microtissues. The co-cultured HRPTEpiC microtissues showed specific uptake of albumin that could be inhibited by cadmium and gentamycin. In conclusion, we established a reliable hanging drop protocol to obtain functional kidney microtissues with proximal tubular epithelial cell lines. These microtissues could be used for high-throughput drug and toxicology screenings, with endocytosis as a functional readout.

Document type Article de périodique (Journal article) – Article de recherche
Access type Accès restreint
Publication date 2016
Language Anglais
Journal information "Pflügers Archiv - European journal of physiology" - Vol. 468, no. 4, p. 739-750 (2016)
Peer reviewed yes
Publisher Springer ((Germany) Berlin)
issn 0031-6768
e-issn 1432-2013
Publication status Publié
Affiliations UCL - SSS/IREC/NEFR - Pôle de Néphrologie
UCL - (SLuc) Service de néphrologie
Keywords Albumin ; Drug toxicity ; Endocytosis ; Epithelial cells ; Primary culture
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Bibliographic reference Prange, Jenny A ; Bieri, Manuela ; Segerer, Stephan ; Burger, Charlotte ; Kaech, Andres ; et. al. Human proximal tubule cells form functional microtissues.. In: Pflügers Archiv - European journal of physiology, Vol. 468, no. 4, p. 739-750 (2016)
Permanent URL http://hdl.handle.net/2078.1/171928