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SELECTBIO Conferences Stem Cells and Antibodies in Drug Discovery Europe 2018

Stem Cells and Antibodies in Drug Discovery Europe 2018 Agenda

Automated Hepatocyte Differentiation From Pluripotent Stem Cells and Their Use in Drug Screening

David Hay, Principal Investigator, MRC Centre for Regenerative Medicine, University of Edinburgh

In both preclinical and clinical testing, drug induced liver injury (DILI) is the major cause of drug attrition, costing pharmaceutical companies millions of dollars every year (Ware & Kehtani 2017, Trends in Biotechnology).  It is therefore essential to rapidly identify and remove drug candidates that pose a risk, decreasing development costs and improving post market success. Currently primary human hepatocytes (PHH) are the most widely used cell type for the prediction of DILI. However when cultured in vitro PHH do not proliferate, rapidly lose their phenotype, in particular drug metabolism. Furthermore, sources are limited and usually isolated from diseased tissue limiting their utility in large scale drug screening. The cost effective delivery of human tissue from a renewable resource, such as pluripotent stem cells (PSCs), offers a possible solution to the issues associated with human somatic cells for in vitro testing. Of note, PSC derived hepatocytes have previously been shown to be as sensitive and predictable as the pharmaceutical ‘gold standard’ PHH (Szkolnicka et al 2014, Stem Cells Translational Medicine and Szkolnicka et al 2016, Stem Cells Translational Medicine). More recently, improvements in culture methods and protocols have led to further improvement in hepatocyte function and stability. The focus of our recent research has been to combine defined differentiation with automation to deliver reliable and stable liver models at scale. In order to multiplex our cell based screening tool we have used a cytological assay that ‘paints the cell’, specifically marking intracellular components of the cell, allowing high through put and multi-parametric analysis (Gustafsdottir et al 2013, PLoS One; Bray et al 2016, Nature Protocols). PSCs were differentiated into hepatocyte like cells (HLCs) using a previously published protocol (Cameron et al 2015, Stem Cell Reports). Once mature HLCs were incubated with a pharmaceutical grade compound library provided by AstraZeneca. Following 48 hours exposure, we scored for drug toxicity, using multiple endpoints and correctly identified drug toxicity in compounds tested. In conclusion, we have developed an automated, reproducible, and scalable platform to generate functional hepatocytes for human drug screening. Importantly, the production method is GLP compliant and cost effective (Cameron et al 2015, Stem Cell Reports). Going forward our automated system will be further miniaturised and fine-tuned to study genetic variation in the human population.