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Updated: Feb 22, 2026
Human Liver Microphysiological System for Assessing Drug-Induced Liver Toxicity In Vitro
Published on: January 31, 2022
Perfused human hepatocyte microtissues identify reactive metabolite-forming and mitochondria-perturbing hepatotoxins
Cliff Rowe1, Mohsen Shaeri1, Emma Large1
1CN Bio Innovations Limited, BioPark, Broadwater Road, Welwyn Garden City AL7 3AX, UK.
Abstract:
Hepatotoxins cause liver damage via many mechanisms but the formation of reactive metabolites and/or damage to liver mitochondria are commonly implicated. We assess 3D human primary hepatocyte microtissues as a platform for hepatotoxicity studies with reactive metabolite-forming and mitochondria-perturbing compounds. We show that microtissues formed from cryopreserved human hepatocytes had bile canaliculi, transcribed mRNA from genes associated with xenobiotic metabolism and expressed functional cytochrome P450 enzymes. Hierarchical clustering was used to distinguish dose-dependent hepatotoxicity elicited by clozapine, fialuridine and acetaminophen (APAP) from control cultures and less liver-damaging compounds, olanzapine and entecavir. The regio-isomer of acetaminophen, N-acetyl-meta-aminophenol (AMAP) clustered with the hepatotoxic compounds. The principal metabolites of APAP were formed and dose-dependent changes in metabolite profile similar to those seen in patient overdose was observed. The toxicological profile of APAP was indistinguishable from that of AMAP, confirming AMAP as a human hepatotoxin. Tissue oxygen consumption rate was significantly decreased within 2h of exposure to APAP or AMAP, concomitant with glutathione depletion. These data highlight the potential utility of perfused metabolically functional human liver microtissues in drug development and mechanistic toxicology.
Insights
3D human liver microtissues effectively model drug-induced liver injury, identifying acetaminophen (APAP) and its isomer (AMAP) as potent hepatotoxins by analyzing metabolite formation and mitochondrial dysfunction.
Area of Science:
- Hepatology
- Toxicology
- Drug Development
Background:
- Hepatotoxins cause liver damage through mechanisms like reactive metabolites and mitochondrial injury.
- 3D human primary hepatocyte microtissues offer a promising in vitro model for studying hepatotoxicity.
Purpose of the Study:
- To assess 3D human primary hepatocyte microtissues for hepatotoxicity studies.
- To investigate the toxicological profiles of reactive metabolite-forming and mitochondria-perturbing compounds.
Main Methods:
- Cultured cryopreserved human hepatocytes into 3D microtissues.
- Analyzed microtissue function including bile canaliculi formation, mRNA expression, and cytochrome P450 activity.
- Utilized hierarchical clustering to differentiate dose-dependent toxicity of compounds like acetaminophen (APAP).
- Measured tissue oxygen consumption and glutathione levels.
Main Results:
- Microtissues exhibited key liver functions: bile canaliculi, xenobiotic metabolism gene expression, and functional cytochrome P450s.
- Hierarchical clustering successfully distinguished hepatotoxic compounds (clozapine, fialuridine, APAP) from less toxic ones.
- The acetaminophen isomer, N-acetyl-meta-aminophenol (AMAP), clustered with hepatotoxic compounds and showed similar toxicity.
- APAP and AMAP exposure rapidly decreased oxygen consumption and depleted glutathione.
Conclusions:
- 3D human liver microtissues are a valuable platform for drug development and mechanistic toxicology.
- AMAP is confirmed as a human hepatotoxin, with a toxicological profile indistinguishable from APAP.
- The microtissue model accurately reflects in vivo observations of APAP overdose, including metabolite profiles and mitochondrial effects.
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