Activities per year
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).
|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.
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- 1 Participation in conference
Philipp Treskes (Participant)9 Apr 2014 → 13 Sep 2014
Activity: Participating in or organising an event types › Participation in conference