Human hepatic HepaRG co-culture model as a sensitive and non-invasive toxicological platform using ECIS (electrical cell-substrate impedance sensing) biosensors

W. Gamal, P. Treskes, C. Chesne, John Plevris, P.-o. Bagnaninchi, L. Nelson

Research output: Contribution to journalArticlepeer-review

Abstract

Background and aims: A robust HTS-compatible multiparametric platform for assessing drug hepatotoxicity would provide a new paradigm for in vitro toxicology. Using the highly-differentiated human hepatic cell line HepaRG as a surrogate to primary human hepatocytes, our aim was to profile real-time changes in cellular impedance, |Z|, in response to paracetamol toxicity. Such dynamic screening allows capture and modeling of multiparametric, high-content data using ECIS biosensors.

Methods: HepaRG cells were cultured to confluence (terminally-differentiated Hepatocyte:Cholangiocyte co-culture) on ECIS-8W10+ and 8W1E-dd 8-well (250,000 cells/well) electrode arrays. Following CYP3A4 induction (day 8), with Rifampicin (24h), a paracetamol time-/dose-response [DMSO-vehicle as controls] was monitored with Impedance |Z| measurements taken every 180 sec over a 500Hz to 64kHz frequency (f) range, for 96h post-induction. Parallel ATP-depletion viability assays were performed (Promega-CellTiter©-Glo). ECIS-8W10+ had 40 electrodes/well (averaging |Z| signal over hundreds of cells). ECIS-8W1E-dd arrays had a range of electrode sizes ranging from 250um down to 20um (sample specific cell populations).
Results
|Z|-time curves showed highly sensitive dose-related response to paracetamol over the critical 24h period [Fig.1], with decreasing |Z| values reflecting disruption of cell-cell/cell-substrate interactions (f=4KHz); whilst at f=64kHz, |Z| changes were indicative of metabolic derangement and loss of membrane integrity. HepaRG ATP content mirrored Z changes. |Z|-spectra modelling (utilizing ECIS algorithms) supported these observations, and provided further evidence of disruption of barrier function/ cell-adhesion parameters.

Conclusions: Real-time high resolution tracking of morphological changes following hepatotoxic challenge was observed. The HepaRG-ECIS platform may offer a breakthrough technology for pre-clinical in vitro hepatotoxicology.
Original languageEnglish
Pages (from-to)S177-S178
JournalJournal of Hepatology
Volume60
Issue number1
DOIs
Publication statusPublished - 1 Apr 2014

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