Biotechnology Letters

, Volume 40, Issue 5, pp 755–763 | Cite as

Hepatic esterase activity is increased in hepatocyte-like cells derived from human embryonic stem cells using a 3D culture system

  • Young-Jun Choi
  • Hyemin Kim
  • Ji-Woo Kim
  • Seokjoo Yoon
  • Han-Jin Park
Original Research Paper



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.


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.


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.


Human embryonic stem cell 3D culture Hepatic maturation Hepatic esterases Drug transporters 



The authors would like to thank the CHA Stem Cell Institute (CHA University, Korea) for providing the CHA-hES15 cell line. This work was funded by the Basic Science Research Program through National Research Foundation (NRF) funding from the Ministry of Science and ICT, Republic of Korea (No. NRF-2017R1C1B2010444).

Compliance with ethical standards

Conflict of interest

The authors declare no conflict of interest.


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© Springer Science+Business Media B.V., part of Springer Nature 2018

Authors and Affiliations

  1. 1.Predictive Model Research CenterKorea Institute of ToxicologyDaejeonRepublic of Korea
  2. 2.Human and Environmental Toxicology, School of EngineeringUniversity of Science and TechnologyDaejeonRepublic of Korea

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