Development of a human liver microphysiological coculture system for higher throughput chemical safety assessment

Blanche C. Ip*, Samantha J. Madnick, Sophia Zheng, Tessa C.A. van Tongeren, Susan J. Hall, Hui Li, Suzanne Martin, Sandrine Spriggs, Paul Carmichael, Wei Chen, David Ames, Lori A. Breitweiser, Heather E. Pence, Andrew J. Bowling, Kamin J. Johnson, Richard Cubberley, Jeffrey R. Morgan, Kim Boekelheide*

*Corresponding author for this work

Research output: Contribution to journalArticleAcademicpeer-review

1 Citation (Scopus)

Abstract

Chemicals in the systemic circulation can undergo hepatic xenobiotic metabolism, generate metabolites, and exhibit altered toxicity compared with their parent compounds. This article describes a 2-chamber liver-organ coculture model in a higher-throughput 96-well format for the determination of toxicity on target tissues in the presence of physiologically relevant human liver metabolism. This 2-chamber system is a hydrogel formed within each well consisting of a central well (target tissue) and an outer ring-shaped trough (human liver tissue). The target tissue chamber can be configured to accommodate a three-dimensional (3D) spheroid-shaped microtissue, or a 2-dimensional (2D) cell monolayer. Culture medium and compounds freely diffuse between the 2 chambers. Human-differentiated HepaRG liver cells are used to form the 3D human liver microtissues, which displayed robust protein expression of liver biomarkers (albumin, asialoglycoprotein receptor, Phase I cytochrome P450 [CYP3A4] enzyme, multidrug resistance-associated protein 2 transporter, and glycogen), and exhibited Phase I/II enzyme activities over the course of 17 days. Histological and ultrastructural analyses confirmed that the HepaRG microtissues presented a differentiated hepatocyte phenotype, including abundant mitochondria, endoplasmic reticulum, and bile canaliculi. Liver microtissue zonation characteristics could be easily modulated by maturation in different media supplements. Furthermore, our proof-of-concept study demonstrated the efficacy of this coculture model in evaluating testosterone-mediated androgen receptor responses in the presence of human liver metabolism. This liver-organ coculture system provides a practical, higher-throughput testing platform for metabolism-dependent bioactivity assessment of drugs/chemicals to better recapitulate the biological effects and potential toxicity of human exposures.
Original languageEnglish
Pages (from-to)227-245
JournalToxicological sciences
Volume199
Issue number2
Early online date9 Feb 2024
DOIs
Publication statusPublished - 2024

Fingerprint

Dive into the research topics of 'Development of a human liver microphysiological coculture system for higher throughput chemical safety assessment'. Together they form a unique fingerprint.

Cite this