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Area of Science:

Hepatology
Drug Discovery
Toxicology

Background:

Rodent models for drug toxicity testing have limitations due to genetic diversity in human populations.
Current in vitro methods using primary mouse hepatocytes (PMHs) have short functional lifespans, hindering long-term compound exposure studies.
The liver is a major organ for drug attrition, necessitating better predictive models for hepatotoxicity.

Purpose of the Study:

To develop a stable in vitro culture system for primary mouse hepatocytes (PMHs) that maintains function for extended periods.
To compare the performance of the novel culture system against existing methods for assessing liver functions and compound responses.
To enable strain- and species-specific comparisons of drug effects using mouse and human hepatocytes.

Main Methods:

Fabrication of mouse micropatterned cocultures (mMPCCs) using PMHs and 3T3-J2 fibroblasts.
Culturing mMPCCs for up to 4 weeks.
Assessing liver functions including albumin secretion, urea synthesis, and cytochrome P450 (CYP) activities.
Exposing mMPCCs to 14 compounds to evaluate CYP induction and hepatotoxicity.
Comparing mMPCC results with primary human hepatocytes (PHHs).

Main Results:

mMPCCs maintained high levels of liver functions for 4 weeks, significantly outperforming standard sandwich cultures.
Albumin secretion, urea synthesis, and CYP activities in mMPCCs were substantially higher than in monocultures.
Functional longevity allowed for in vivo relevant comparisons of strain-specific responses to compounds.
mMPCCs facilitated comparisons of drug responses between mouse and human hepatocytes.

Conclusions:

Mouse micropatterned cocultures (mMPCCs) offer a robust platform for long-term liver cell culture, preserving key liver functions.
mMPCCs enable the assessment of strain- and species-specific drug effects, aiding in the prediction of human hepatotoxicity.
This model can leverage mouse genetics to identify susceptible subpopulations and understand genetic determinants of severe drug-induced liver injury.