Accurate in vitro pharmacological profiling at an early stage is critical for lead optimization. Combining live content and high-content imaging, Porsolt has developed a cost-efficient strategy to accurately determine the efficacy and toxicity profile of lead coumpounds, and explore their underlying mechanism of action.
Phalloidin and MRP2 immunofluorescence reveals the bile canaliculi network formed by polarized primary rat hepatocytes.
To accurately characterize the effects and mechanism of action of drug candidates, we select physiologically relevant cellular models, which recapitulate key functional and molecular features:
> Cell lines, primary cells or differentiated stem cells in monolayer cultures (mono- or multi-cell type cultures)
> Polarized primary cell cultures
> 3D spheroid monocell type cultures
> In vitro disease models (pharmacological treatment of genetic modification)
Our drug profiling strategy starts with the kinetic monitoring of compounds' effects over time in live cells.
Live content imaging enables the multiplexed analysis of phenotypic features such as cell proliferation, cell death, neurite outgrowth, etc., over several days:
> Cost-efficient profiling: a single well provides the whole kinetic data of a given condition
> Determination of optimal exposure duration and drug concentration, through time-dependent EC-50 values
Cellular markers, out of a panel of 60 molecular assays, are selected and analyzed by high-content imaging to investigate the effects of the compound on specific signaling pathways:
> Accurate: the expression and intracellular localization of specific markers are quantified at the single cell level, allowing subpopulations analysis
> Robust: molecular markers are validated with specific reference compounds
In addition to in vitro mechanistic studies, Porsolt offers to validate the mechanism of action of lead compounds in vivo, through the multiplexed quantification of biomarkers in blood or tissue samples by high-content analysis.