Hepatic esterase activity is increased in hepatocyte-like cells derived from human embryonic stem cells using a 3D culture system
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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.
KeywordsHuman 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.
- Davies BE (2010) Pharmacokinetics of oseltamivir: an oral antiviral for the treatment and prophylaxis of influenza in diverse populations. J Antimicrob Chemother 65(Suppl 2):5–10Google Scholar
- Godoy P, Hewitt NJ, Albrecht U, Andersen ME et al (2013) Recent advances in 2D and 3D in vitro systems using primary hepatocytes, alternative hepatocyte sources and non-parenchymal liver cells and their use in investigating mechanisms of hepatotoxicity, cell signaling and ADME. Arch Toxicol 87:1315–1530CrossRefPubMedCentralPubMedGoogle Scholar
- Nagamoto Y, Tashiro K, Takayama K, Ohashi K, Kawabata K, Sakurai F, Tachibana M, Hayakawa T, Furue MK, Mizuguchi H (2012) The promotion of hepatic maturation of human pluripotent stem cells in 3D co-culture using type I collagen and Swiss 3T3 cell sheets. Biomaterials 33:4526–4534CrossRefPubMedGoogle Scholar
- Schyschka L, Sánchez JJM, Wang Z, Burkhardt B, Müller-Vieira U, Zeilinger K, Bachmann A, Nadalin S, Damm G, Nussler AK (2013) Hepatic 3D cultures but not 2D cultures preserve specific transporter activity for acetaminophen-induced hepatotoxicity. Arch Toxicol 87:1581–1593CrossRefPubMedGoogle Scholar
- Shi D, Yang J, Yang D, LeCluyse EL, Black C, You L, Akhlaghi F, Yan B (2006) Anti-influenza prodrug oseltamivir is activated by carboxylesterase human carboxylesterase 1, and the activation is inhibited by antiplatelet agent clopidogrel. J Pharmacol Exp Ther 319:1477–1484CrossRefPubMedGoogle Scholar