| Literature DB >> 27479365 |
Susanne Ramm1,2, Melanie Adler1,2, Vishal S Vaidya1,2,3.
Abstract
Kidney toxicity due to drugs and chemicals poses a significant health burden for patients and a financial risk for pharmaceutical companies. However, currently no sensitive and high-throughput in vitro method exists for predictive nephrotoxicity assessment. Primary human proximal tubular epithelial cells (HPTECs) possess characteristics of differentiated epithelial cells, making them a desirable model to use in in vitro screening systems. Additionally, heme oxygenase 1 (HO-1) protein expression is upregulated as a protective mechanism during kidney toxicant-induced oxidative stress or inflammation in HPTECs and can therefore be used as a biomarker for nephrotoxicity. In this article, we describe two different methods to screen for HO-1 increase: A homogeneous time resolved fluorescence (HTRF) assay and an immunofluorescence assay. The latter provides lower throughput but higher sensitivity due to the combination of two readouts, HO-1 intensity and cell number. The methods described in the protocol are amendable for other cell types as well. © 2016 by John Wiley & Sons, Inc.Entities:
Keywords: HTRF; biomarker; heme oxygenase 1; high-throughput; in vitro; primary human proximal tubular epithelial cells; screening
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Year: 2016 PMID： 27479365 PMCID： PMC4981919 DOI： 10.1002/cptx.12
Source DB: PubMed Journal: Curr Protoc Toxicol ISSN： 1934-9254