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Cited by 1183D Printing in Biomedical ResearchDrug-Induced Hepatotoxicity and ProtectionLiver physiology and pathology

3D spheroid cultures improve the metabolic gene expression profiles of HepaRG cells

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Takahashi, Yu, Hori, Yuji, Yamamoto, Tomohisa, Urashima, Toshiki, Ohara, Yasunori, Tanaka, Hideo·2015·Bioscience Reports
DOI 10.1042/bsr20150034PMC PMC4613666
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Abstract

3D (three-dimensional) cultures are considered to be an effective method for toxicological studies; however, little evidence has been reported whether 3D cultures have an impact on hepatocellular physiology regarding lipid or glucose metabolism. In the present study, we conducted physiological characterization of hepatoma cell lines HepG2 and HepaRG cells cultured in 3D conditions using a hanging drop method to verify the effect of culture environment on cellular responses. Apo (Apolipoprotein)B as well as albumin secretion was augmented by 3D cultures. Expression of genes related to not only drug, but also glucose and lipid metabolism were significantly enhanced in 3D cultured HepaRG spheroids. Furthermore, mRNA levels of CYP (cytochrome P450) enzymes following exposure to corresponding inducers increased under the 3D condition. These data suggest that this simple 3D culture system without any special biomaterials can improve liver-specific characteristics including lipid metabolism. Considering that the system enables high-throughput assay, it may become a powerful tool for compound screening concerning hepatocellular responses in order to identify potential drugs.

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  • In a number of studies, 3D HepG2 spheroids demonstrated increased secretion of albumin and apolipoproteins, as well as enhanced metabolic functions compared to 2D monocultures.

    “In a number of studies, 3D HepG2 spheroids demonstrated increased secretion of albumin and apolipoproteins, as well as enhanced metabolic functions compared to 2D monocultures [169,170].”

    From The HepG2 Cell Line as a Model for Studying Metabolic Dysfunction-Associated Steatotic Liver Disease (2026) · cites this paper as [169]

    The Results sections on albumin and apoB secretion report higher secretion per viable cell in 3D HepG2 spheroids than in 2D cultures. But the Results section on liver-specific gene expression says metabolic gene expression in HepG2 remained almost unchanged; the reported improvements were in HepaRG cells. The paper therefore does not establish enhanced metabolic functions in HepG2 beyond the measured secretion outcomes. (LLM, conf=0.89)

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Research works printed but not linked: 28

  1. [2]Zheng W. Thorne N. McKew J.C. Phenotypic screens as a renewed approach for drug discovery Drug Discov. Today 2013 18 1067 1073 10.1016/j.drudis.2013.07.001 23850704 PMC4531371 · DOI
  2. [3]Pampaloni F. Stelzer E.H. Masotti A. Three-dimensional tissue models for drug discovery and toxicology Recent Pat. Biotechnol. 2009 3 103 117 10.2174/187220809788700201 19519566 · DOI
  3. [4]Meng Q. Three-dimensional culture of hepatocytes for prediction of drug-induced hepatotoxicity Expert Opin. Drug Metab. Toxicol. 2010 6 733 746 10.1517/17425251003674356 20380484 · DOI
  4. [5]Rimann M. Graf-Hausner U. Synthetic 3D multicellular systems for drug development Curr. Opin. Biotechnol. 2012 23 803 809 10.1016/j.copbio.2012.01.011 22326911 · DOI
  5. [6]Ramaiahgari S.C. den Braver M.W. Herpers B. Terpstra V. Commandeur J.N. van de Water B. Price L.S. A 3D in vitro model of differentiated HepG2 cell spheroids with improved liver-like properties for repeated dose high-throughput toxicity studies Arch. Toxicol. 2014 88 1083 1095 24599296
  6. [7]Tung Y.C. Hsiao A.Y. Allen S.G. Torisawa Y.S. Ho M. Takayama S. High-throughput 3D spheroid culture and drug testing using a 384 hanging drop array Analyst 2011 136 473 478 10.1039/C0AN00609B 20967331 PMC7454010 · DOI · Paper page
  7. [8]Karlsson H. Fryknas M. Larsson R. Nygren P. Loss of cancer drug activity in colon cancer HCT-116 cells during spheroid formation in a new 3-D spheroid cell culture system Exp. Cell Res. 2012 318 1577 1585 10.1016/j.yexcr.2012.03.026 22487097 · DOI · Paper page
  8. [9]Mueller D. Kramer L. Hoffmann E. Klein S. Noor F. 3D organotypic HepaRG cultures as in vitro model for acute and repeated dose toxicity studies Toxicol. In Vitro 2014 28 104 112 10.1016/j.tiv.2013.06.024 23850736 · DOI
  9. [10]Chen L. Xiao Z. Meng Y. Zhao Y. Han J. Su G. Chen B. Dai J. The enhancement of cancer stem cell properties of MCF-7 cells in 3D collagen scaffolds for modeling of cancer and anti-cancer drugs Biomaterials 2012 33 1437 1444 10.1016/j.biomaterials.2011.10.056 22078807 · DOI
  10. [11]Pescosolido L. Schuurman W. Malda J. Matricardi P. Alhaique F. Coviello T. van Weeren P.R. Dhert W.J. Hennink W.E. Vermonden T. Hyaluronic acid and dextran-based semi-IPN hydrogels as biomaterials for bioprinting Biomacromolecules 2011 12 1831 1838 10.1021/bm200178w 21425854 · DOI
  11. [12]Trojani C. Weiss P. Michiels J.F. Vinatier C. Guicheux J. Daculsi G. Gaudray P. Carle G.F. Rochet N. Three-dimensional culture and differentiation of human osteogenic cells in an injectable hydroxypropylmethylcellulose hydrogel Biomaterials 2005 26 5509 5517 10.1016/j.biomaterials.2005.02.001 15860207 · DOI
  12. [13]Wang X. Ito Y. Luan X. Yamane A. Diekwisch T.G. Amelogenin sequence and enamel biomineralization in Rana pipiens J. Exp. Zool. B Mol. Dev. Evol. 2005 304 177 186 10.1002/jez.b.21035 15744754 · DOI
  13. [14]Kelm J.M. Fussenegger M. Microscale tissue engineering using gravity-enforced cell assembly Trends Biotechnol. 2004 22 195 202 10.1016/j.tibtech.2004.02.002 15038925 · DOI
  14. [15]Fayad W. Rickardson L. Haglund C. Olofsson M.H. D'Arcy P. Larsson R. Linder S. Fryknas M. Identification of agents that induce apoptosis of multicellular tumour spheroids: enrichment for mitotic inhibitors with hydrophobic properties Chem. Biol. Drug Des. 2011 78 547 557 10.1111/j.1747-0285.2011.01170.x 21726416 · DOI
  15. [16]Godoy P. Hengstler J.G. Ilkavets I. Meyer C. Bachmann A. Muller A. Tuschl G. Mueller S.O. Dooley S. Extracellular matrix modulates sensitivity of hepatocytes to fibroblastoid dedifferentiation and transforming growth factor beta-induced apoptosis Hepatology 2009 49 2031 2043 10.1002/hep.22880 19274752 · DOI
  16. [17]Aninat C. Piton A. Glaise D. Le Charpentier T. Langouet S. Morel F. Guguen-Guillouzo C. Guillouzo A. Expression of cytochromes P450, conjugating enzymes and nuclear receptors in human hepatoma HepaRG cells Drug Metab. Dispos. 2006 34 75 83 10.1124/dmd.105.006759 16204462 · DOI
  17. [18]Lubberstedt M. Muller-Vieira U. Mayer M. Biemel K.M. Knospel F. Knobeloch D. Nussler A.K. Gerlach J.C. Zeilinger K. HepaRG human hepatic cell line utility as a surrogate for primary human hepatocytes in drug metabolism assessment in vitro J. Pharmacol. Toxicol. Methods 2011 63 59 68 10.1016/j.vascn.2010.04.013 20460162 · DOI
  18. [19]Andersson T.B. Kanebratt K.P. Kenna J.G. The HepaRG cell line: a unique in vitro tool for understanding drug metabolism and toxicology in human Expert Opin. Drug Metab. Toxicol. 2012 8 909 920 10.1517/17425255.2012.685159 22568886 · DOI
  19. [20]Wilkening S. Stahl F. Bader A. Comparison of primary human hepatocytes and hepatoma cell line Hepg2 with regard to their biotransformation properties Drug Metab. Dispos. 2003 31 1035 1042 10.1124/dmd.31.8.1035 12867492 · DOI · Paper page
  20. [21]Leite S.B. Wilk-Zasadna I. Zaldivar J.M. Airola E. Reis-Fernandes M.A. Mennecozzi M. Guguen-Guillouzo C. Chesne C. Guillou C. Alves P.M. Coecke S. Three-dimensional HepaRG model as an attractive tool for toxicity testing Toxicol. Sci. 2012 130 106 116 10.1093/toxsci/kfs232 22843569 · DOI
  21. [22]Takayama K. Kawabata K. Nagamoto Y. Kishimoto K. Tashiro K. Sakurai F. Tachibana M. Kanda K. Hayakawa T. Furue M.K. Mizuguchi H. 3D spheroid culture of hESC/hiPSC-derived hepatocyte-like cells for drug toxicity testing Biomaterials 2013 34 1781 1789 10.1016/j.biomaterials.2012.11.029 23228427 · DOI
  22. [23]Vukicevic S. Kleinman H.K. Luyten F.P. Roberts A.B. Roche N.S. Reddi A.H. Identification of multiple active growth factors in basement membrane Matrigel suggests caution in interpretation of cellular activity related to extracellular matrix components Exp. Cell Res. 1992 202 1 8 10.1016/0014-4827(92)90397-Q 1511725 · DOI · Paper page
  23. [24]Lehmann J.M. McKee D.D. Watson M.A. Willson T.M. Moore J.T. Kliewer S.A. The human orphan nuclear receptor PXR is activated by compounds that regulate CYP3A4 gene expression and cause drug interactions J. Clin. Invest. 1998 102 1016 1023 10.1172/JCI3703 9727070 PMC508967 · DOI
  24. [25]Daitoku H. Yamagata K. Matsuzaki H. Hatta M. Fukamizu A. Regulation of PGC-1 promoter activity by protein kinase B and the forkhead transcription factor FKHR Diabetes 2003 52 642 649 10.2337/diabetes.52.3.642 12606503 · DOI · Paper page
  25. [26]Kawaguchi T. Takenoshita M. Kabashima T. Uyeda K. Glucose and cAMP regulate the L-type pyruvate kinase gene by phosphorylation/dephosphorylation of the carbohydrate response element binding protein Proc. Natl. Acad. Sci. U.S.A. 2001 98 13710 13715 10.1073/pnas.231370798 11698644 PMC61106 · DOI
  26. [27]Takahashi Y. Shinoda A. Furuya N. Harada E. Arimura N. Ichi I. Fujiwara Y. Inoue J. Sato R. Perilipin-mediated lipid droplet formation in adipocytes promotes sterol regulatory element-binding protein-1 processing and triacylglyceride accumulation PLoS One 2013 8 e64605 10.1371/journal.pone.0064605 23734208 PMC3667186 · DOI
  27. [28]Ananthanarayanan M. Balasubramanian N. Makishima M. Mangelsdorf D.J. Suchy F.J. Human bile salt export pump promoter is transactivated by the farnesoid X receptor/bile acid receptor J. Biol. Chem. 2001 276 28857 28865 10.1074/jbc.M011610200 11387316 · DOI
  28. [29]Delerive P. Galardi C.M. Bisi J.E. Nicodeme E. Goodwin B. Identification of liver receptor homolog-1 as a novel regulator of apolipoprotein AI gene transcription Mol. Endocrinol. 2004 18 2378 2387 10.1210/me.2004-0132 15218078 · DOI

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Authors
Takahashi, Y., Hori, Y., Yamamoto, T., Urashima, T., Ohara, Y., & Tanaka, H.
Affiliations
Japan Tobacco (Japan) (JP)
Corresponding author
  • Takahashi, Yu
  • Hori, Yuji
  • Yamamoto, Tomohisa
  • Urashima, Toshiki
  • Ohara, Yasunori
  • Tanaka, Hideo
ORCID
Takahashi, Yu
Venue
Bioscience Reports
Publisher
Portland Press Ltd.
Published
2015-06-01
Volume
35
Issue
3
ISSN
  • 0144-8463 (ISSN-L)
  • 1573-4935
DOI
10.1042/bsr20150034
PMC
PMC4613666
OpenAlex
W2284842265
Cited by
118
References
1
Citations by year
2026: 7 · 2025: 7 · 2024: 9 · 2023: 10 · 2022: 11 · 2021: 13 · 2020: 15 · 2019: 10 · 2018: 15 · 2017: 10 · 2016: 7 · 2015: 4
FWCI
5.97
Type
journal article
Language
en
Open access
Licence
Locations
  • Bioscience Reports (publishedVersion, cc-by)
  • Europe PMC (PubMed Central) (publishedVersion, cc-by)
  • PubMed Central (publishedVersion, cc-by)
  • PubMed (publishedVersion)
Topics
  • Physical Sciences › Engineering › Biomedical Engineering › 3D Printing in Biomedical Research
  • Life Sciences › Pharmacology, Toxicology and Pharmaceutics › Pharmacology › Drug-Induced Hepatotoxicity and Protection
  • Health Sciences › Medicine › Hepatology › Liver physiology and pathology
Keywords
3D spheroid culture, HepaRG cells, HepG2 cells, hanging drop culture, lipid metabolism, glucose metabolism, apolipoprotein B, albumin secretion, cytochrome P450 induction, drug screening, in vitro liver models
MeSH terms
Albumins, Apolipoproteins B, Cytochrome P-450 Enzyme System, Glucose, Humans, Lipoproteins, LDL, Liver, Omeprazole, Phenobarbital, Rifampin, Spheroids, Cellular, Cell Culture Techniques, Lipid Metabolism, Hep G2 Cells, High-Throughput Screening Assays, Transcriptome, Cytochrome P-450 Enzyme Inducers

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