Induction of in vitro Metabolic Zonation in Primary Hepatocytes Requires Both Near-Physiological Oxygen Concentration and Flux

Benedikt Scheidecker; Marie Shinohara; Masahiro Sugimoto; Mathieu Danoy; Masaki Nishikawa; Yasuyuki Sakai

doi:10.3389/fbioe.2020.00524

Induction of in vitro Metabolic Zonation in Primary Hepatocytes Requires Both Near-Physiological Oxygen Concentration and Flux

Front Bioeng Biotechnol. 2020 Jun 3:8:524. doi: 10.3389/fbioe.2020.00524. eCollection 2020.

Authors

Benedikt Scheidecker¹, Marie Shinohara², Masahiro Sugimoto³, Mathieu Danoy⁴, Masaki Nishikawa¹, Yasuyuki Sakai¹

Affiliations

¹ Department of Chemical System Engineering, University of Tokyo, Tokyo, Japan.
² Department of Mechanical and Biofunctional Systems, Institute of Industrial Science, The University of Tokyo, Tokyo, Japan.
³ Institute for Advanced Biosciences, Keio University, Yamagata, Japan.
⁴ CNRS UMI 2820, LIMMS, University of Tokyo, Tokyo, Japan.

Abstract

Pre-clinical drug screening is an important step in assessing the metabolic effects and hepatic toxicity of new pharmaceutical compounds. However, due to the complexity of the liver microarchitecture, simplified in vitro models do not adequately reflect in vivo situations. Especially spatial heterogeneity, known as metabolic zonation, is often lost due to limitations introduced by typical culture conditions. By culturing primary rat hepatocytes in varied ambient oxygen levels on either gas-permeable or non-permeable culture plates, we highlight the importance of biomimetic oxygen supply for the targeted induction of zonation-like phenotypes. Resulting cellular profiles illustrate the effect of pericellular oxygen concentration and consumption rates on hepatic functionality in terms of zone-specific metabolism and β-catenin signaling. We show that modulation of ambient oxygen tension can partially induce metabolic zonation in vitro when considering high supply rates, leading to in vivo-like drug metabolism. However, when oxygen supply is limited, similar modulation instead triggers an ischemic reprogramming, resembling metabolic profiles of hepatocellular carcinoma and increasing susceptibility toward drug-induced injury. Application of this knowledge will allow for the development of more accurate drug screening models to better identify adverse effects in hepatic drug metabolism.

Keywords: ADME-Tox; PDMS; hepatocyte; metabolic reprogramming; oxygen supply; zonation.