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Cited by 703D Printing in Biomedical ResearchDrug-Induced Hepatotoxicity and ProtectionAnimal testing and alternatives

Characterization of In Vitro 3D Cell Model Developed from Human Hepatocellular Carcinoma (HepG2) Cell Line

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Štampar, Martina, Breznik, Barbara, Filipič, Metka, Žegura, Bojana·2020·Cells
DOI 10.3390/cells9122557PMC PMC7759933
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Abstract

In genetic toxicology, there is a trend against the increased use of in vivo models as highlighted by the 3R strategy, thus encouraging the development and implementation of alternative models. Two-dimensional (2D) hepatic cell models, which are generally used for studying the adverse effects of chemicals and consumer products, are prone to giving misleading results. On the other hand, newly developed hepatic three-dimensional (3D) cell models provide an attractive alternative, which, due to improved cell interactions and a higher level of liver-specific functions, including metabolic enzymes, reflect in vivo conditions more accurately. We developed an in vitro 3D cell model from the human hepatocellular carcinoma (HepG2) cell line. The spheroids were cultured under static conditions and characterised by monitoring their growth, morphology, and cell viability during the time of cultivation. A time-dependent suppression of cell division was observed. Cell cycle analysis showed time-dependent accumulation of cells in the G0/G1 phase. Moreover, time-dependent downregulation of proliferation markers was shown at the mRNA level. Genes encoding hepatic markers, metabolic phase I/II enzymes, were time-dependently deregulated compared to monolayers. New knowledge on the characteristics of the 3D cell model is of great importance for its further development and application in the safety assessment of chemicals, food products, and complex mixtures.

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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 [170]

    Section 3.4.2 reports higher albumin expression in HepG2 spheroids than in 2D cultures, and section 3.4.3 reports higher expression of metabolic-enzyme genes, describing the spheroids as more metabolically competent. The paper does not establish increased secretion of albumin or apolipoproteins compared with 2D cultures; its own measurements were of gene expression, and its mention of albumin secretion does not make that comparison. Page numbers are unavailable in our copy. (LLM, conf=0.86)

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Cited by

Sorted by citations

Authors
Štampar, M., Breznik, B., Filipič, M., & Žegura, B.
Affiliations
  • Jožef Stefan International Postgraduate School (SI)
  • National Institute of Biology (SI)
Corresponding author
Žegura, Bojana
ORCID
Venue
Cells
Publisher
MDPI AG
Published
2020-11-28
Volume
9
Issue
12
Pages
2557
ISSN
2073-4409 (ISSN-L)
DOI
10.3390/cells9122557
PMC
PMC7759933
OpenAlex
W3106917159
Cited by
70
References
2
Citations by year
2026: 14 · 2025: 18 · 2024: 13 · 2023: 11 · 2022: 11 · 2021: 3
FWCI
1.96
Type
journal article
Language
en
Open access
Licence
Locations
  • Cells (publishedVersion, cc-by)
  • PubMed Central (publishedVersion, cc-by)
  • PubMed (publishedVersion)
  • DiRROS repository (University of Maribor) (publishedVersion, cc-by)
  • DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion, cc-by-sa)
Funders
  • Javna agencija za raziskovalno dejavnost (J1-2465, P1-0245, MR-MStampar, Z3-1870-BBreznik)
  • European Cooperation in Science and Technology (CA16119 (In vitro 3-D total cell guidance and fitness))
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 › Veterinary › Small Animals › Animal testing and alternatives
Keywords
3D cell models, HepG2 cells, hepatocellular carcinoma, liver spheroids, 3Rs, cell cycle arrest, cell proliferation markers, phase I and phase II enzymes, hepatic gene expression, chemical safety assessment, in vitro liver models
MeSH terms
Cell Survival, Carcinoma, Hepatocellular, Humans, Liver, Liver Neoplasms, Down-Regulation, Spheroids, Cellular, Cell Culture Techniques, Hepatocytes, Cell Line, Tumor, Cell Proliferation, Hep G2 Cells

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