Micropatterned Primary Hepatocyte Co-Cultures for Drug Metabolism and Toxicity Studies
Primary hepatocytes display a precipitous decline in phenotypic functions when cultured in a sandwich of extracellular
matrix proteins (i.e. collagen, Matrigel). We describe a human liver model, HepatoPac TM, with precise microscale cytoarchitecture
and optimal stromal interactions (micropatterned co-cultures) that displays stable functions for several weeks
in vitro. Micropatterned co-cultures were coupled with miniaturization strategies (i.e. 24- and 96-well format) and
optimized for the screening of genotype-specific and clinically-relevant drug disposition. DILI can also be assessed using
standard end-points (i.e. ATP depletion, mitochondrial activity). We have investigated the toxicity of several hepatotoxins
(i.e. Trazodone, Isoniazid, Imipramine etc. ) in our model under both acute and chronic dosing regimens and show
concordance with preclinical and clinical findings. Since metabolism is an important determinant to the overall disposition
of drugs and the profile of metabolites can have an impact on efficacy and safety, the utility of micropatterned co-cultures
for prediction of compound clearance and generation of human metabolites was evaluated. Micropatterned co-cultures
classifies compounds based on rates of clearance and generated metabolites arising from both phase 1 and 2 reactions
(single and sequential). Transporter assays (uptake and efflux) could be performed on micropatterned co-cultures towards
simultaneous assessment of the interplay of drug transport, metabolism, and toxicity. Long-term drug-drug interaction
studies (i.e. enzyme induction and inhibition) have also been explored and results indicate that micropatterned co-cultures
are able to recapitulate clinical outcomes. In the future, miniaturized micropatterned co-cultures may find utility in the
development of several classes of therapeutic compounds (drugs, biologics), in evaluating the injury potential of
environmental toxicants, in fundamental investigations of liver physiology, and in personalized medicine for liver disease.
matrix proteins (i.e. collagen, Matrigel). We describe a human liver model, HepatoPac TM, with precise microscale cytoarchitecture
and optimal stromal interactions (micropatterned co-cultures) that displays stable functions for several weeks
in vitro. Micropatterned co-cultures were coupled with miniaturization strategies (i.e. 24- and 96-well format) and
optimized for the screening of genotype-specific and clinically-relevant drug disposition. DILI can also be assessed using
standard end-points (i.e. ATP depletion, mitochondrial activity). We have investigated the toxicity of several hepatotoxins
(i.e. Trazodone, Isoniazid, Imipramine etc. ) in our model under both acute and chronic dosing regimens and show
concordance with preclinical and clinical findings. Since metabolism is an important determinant to the overall disposition
of drugs and the profile of metabolites can have an impact on efficacy and safety, the utility of micropatterned co-cultures
for prediction of compound clearance and generation of human metabolites was evaluated. Micropatterned co-cultures
classifies compounds based on rates of clearance and generated metabolites arising from both phase 1 and 2 reactions
(single and sequential). Transporter assays (uptake and efflux) could be performed on micropatterned co-cultures towards
simultaneous assessment of the interplay of drug transport, metabolism, and toxicity. Long-term drug-drug interaction
studies (i.e. enzyme induction and inhibition) have also been explored and results indicate that micropatterned co-cultures
are able to recapitulate clinical outcomes. In the future, miniaturized micropatterned co-cultures may find utility in the
development of several classes of therapeutic compounds (drugs, biologics), in evaluating the injury potential of
environmental toxicants, in fundamental investigations of liver physiology, and in personalized medicine for liver disease.
