Young-Jun Choi1,2, Hyemin Kim1, Ji-Woo Kim1, Seokjoo Yoon1,2, Han-Jin Park3. 1. Predictive Model Research Center, Korea Institute of Toxicology, Daejeon, 34114, Republic of Korea. 2. Human and Environmental Toxicology, School of Engineering, University of Science and Technology, Daejeon, 34113, Republic of Korea. 3. Predictive Model Research Center, Korea Institute of Toxicology, Daejeon, 34114, Republic of Korea. hjpark@kitox.re.kr.
Abstract
OBJECTIVES: The aim of the study is to generate a spherical three-dimensional (3D) aggregate of hepatocyte-like cells (HLCs) differentiated from human embryonic stem cells and to investigate the effect of the 3D environment on hepatic maturation and drug metabolism. RESULTS: Quantitative real-time PCR analysis indicated that gene expression of mature hepatocyte markers, drug-metabolizing enzymes, and hepatic transporters was significantly higher in HLCs cultured in the 3D system than in those cultured in a two-dimensional system (p < 0.001). Moreover, hepatocyte-specific functions, including albumin secretion and bile canaliculi formation, were increased in HLCs cultured in the 3D system. In particular, 3D spheroidal culture increased expression of CES1 and BCHE, which encode hepatic esterases (p < 0.001). The enhanced activities of these hepatic esterases were confirmed by the cholinesterase activity assay and the increased susceptibility of HLCs to oseltamivir, which is metabolized by CES1. CONCLUSIONS: 3D spheroidal culture enhances the maturation and drug metabolism of stem cell-derived HLCs, and this may help to optimize hepatic differentiation protocols for hepatotoxicity testing.
OBJECTIVES: The aim of the study is to generate a spherical three-dimensional (3D) aggregate of hepatocyte-like cells (HLCs) differentiated from human embryonic stem cells and to investigate the effect of the 3D environment on hepatic maturation and drug metabolism. RESULTS: Quantitative real-time PCR analysis indicated that gene expression of mature hepatocyte markers, drug-metabolizing enzymes, and hepatic transporters was significantly higher in HLCs cultured in the 3D system than in those cultured in a two-dimensional system (p < 0.001). Moreover, hepatocyte-specific functions, including albumin secretion and bile canaliculi formation, were increased in HLCs cultured in the 3D system. In particular, 3D spheroidal culture increased expression of CES1 and BCHE, which encode hepatic esterases (p < 0.001). The enhanced activities of these hepatic esterases were confirmed by the cholinesterase activity assay and the increased susceptibility of HLCs to oseltamivir, which is metabolized by CES1. CONCLUSIONS: 3D spheroidal culture enhances the maturation and drug metabolism of stem cell-derived HLCs, and this may help to optimize hepatic differentiation protocols for hepatotoxicity testing.
Entities:
Keywords:
3D culture; Drug transporters; Hepatic esterases; Hepatic maturation; Human embryonic stem cell
Authors: Mania Ackermann; Kathrin Haake; Henning Kempf; Paul Kaschutnig; Anna-Carina Weiss; Ariane H H Nguyen; Markus Abeln; Sylvia Merkert; Mark Phillip Kühnel; Dorothee Hartmann; Danny Jonigk; Thomas Thum; Andreas Kispert; Michael D Milsom; Nico Lachmann Journal: Haematologica Date: 2021-05-01 Impact factor: 11.047