Three-Dimensional Dynamic Cell Models for Metabolic Dysfunction-Associated Steatotic Liver Disease Progression
Open accessAbstract
Objective: Metabolic dysfunction-associated steatotic liver disease (MASLD) is a complex, progressive disorder involving multiple cell types, ranging from simple steatosis to metabolic dysfunction-associated steatohepatitis (MASH), characterized by pro-inflammatory macrophage activation, and can eventually advance to fibrosis, initiated by hepatic stellate cells (HSCs). In vitro multi-cell coculture models are vital tools for elucidating the mechanisms underlying MASLD. Impact Statement: Existing in vitro models for MASLD, including traditional 2-dimensional (2D) cultures and advanced organ-on-a-chip and organoid systems, face challenges in representing multiple cell types and analyzing them individually. Here, utilizing a cell carrier developed in our laboratory, we introduce a series of 3D dynamic coculture models that simulate different stages of MASLD progression and enable individual cell type analysis. Introduction: Currently, no single system provides an optimal balance of control, reproducibility, and analytical convenience. Most in vitro models lack the ability to isolate and analyze individual cell types post-culture, making it difficult to study cell-specific responses in MASLD progression. Methods: The 3D hollow porous sphere cell carrier allows cells to grow on its surface, while the culture device (mini-bioreactor) creates a dynamic environment. The 3 distinct MASLD models were established based on cocultured cell types: steatosis (hepatocytes only), MASH (hepatocytes and macrophages in a 4:1 ratio), and fibrosis (hepatocytes, macrophages, and HSCs in an 8:2:1 ratio). Well-established MASLD mouse models were employed to validate our in vitro 3D dynamic MASLD models, using 7-week-old male C57BL/6J mice fed a high-fat diet. Results: Our models demonstrate a progressive decline in hepatocyte viability and increased lipid accumulation, mirroring in vivo pathology. Additionally, gene expression profiles of our models align with those observed in MASLD-affected mouse livers. Notably, comparative analysis highlights the role of pro-inflammatory macrophages in disrupting hepatocyte lipid metabolism. Conclusion: These models offer a robust platform for investigating MASLD mechanisms and show potential for screening anti-MASLD therapeutics.
The claims and statistics of this paper haven’t been checked yet.
Run the check — free, no account needed. It reads the open-access text and checks the sources it cites.
Integrity signals
Registry
Cites
Statistics
Claims the paper makes
The claims and statistics of this paper haven’t been checked yet.
Run the check — free, no account needed. It reads the open-access text and checks the sources it cites.
Claims other papers make about this one
Each is checked against this paper’s own text: does it say what the citing paper says it does?
For tasks related to disease progression (inflammation, fibrosis), as well as for rigorous studies of drug metabolism and toxicity, HepG2 should be used with caution, preferably as part of more complex systems (co-cultures, 3D platforms) or in combination with alternative models (HepaRG, primary hepatocytes).
“For tasks related to disease progression (inflammation, fibrosis), as well as for rigorous studies of drug metabolism and toxicity, HepG2 should be used with caution, preferably as part of more complex systems (co-cultures, 3D platforms) or in combination with alternative models (HepaRG, primary hepatocytes) [158,159,160,161].”
From The HepG2 Cell Line as a Model for Studying Metabolic Dysfunction-Associated Steatotic Liver Disease (2026) · cites this paper as [160]
Split into 2 atoms — 0 confirmed, 2 partially supported, 0 disputed, 0 misattributed, 0 unverifiable. Expand the claim to see per-atom verdicts.
Experimental — accuracy not yet measured
Self-check
Percentages, sums and reported differences re-computed from the paper’s own numbers.
The self-check needs the paper’s full text.
Cites
Mechanisms of NAFLD development and therapeutic strategies
Friedman, S. L., Neuschwander-Tetri, B. A., Rinella, M., & Sanyal, A. J. (2018) · Nature Medicine · doi:10.1038/s41591-018-0104-9 · Cited by 4,354 · see citing works
Donnelly, K. L., Smith, C. I., Schwarzenberg, S. J., Jessurun, J., Boldt, M. D., & Parks, E. J. (2005) · Journal of Clinical Investigation · doi:10.1172/jci23621 · Cited by 3,604 · see citing works
Younossi, Z. M., Golabi, P., Paik, J. M., Henry, A., Van Dongen, C., & Henry, L. (2023) · Hepatology · doi:10.1097/hep.0000000000000004 · Cited by 3,383 · see citing works
Modeling Steatohepatitis in Humans with Pluripotent Stem Cell-Derived Organoids
Ouchi, R., Togo, S., Kimura, M., Shinozawa, T., Koido, M., Koike, H., Thompson, W., Karns, R. A., Mayhew, C. N., McGrath, P. S., et al. (2019) · Cell Metabolism · doi:10.1016/j.cmet.2019.05.007 · Cited by 537 · see citing works
Understanding lipotoxicity in NAFLD pathogenesis: is CD36 a key driver?
Rada, P., González‐Rodríguez, Á., García‐Monzón, C., & Valverde, Á. M. (2020) · Cell Death and Disease · doi:10.1038/s41419-020-03003-w · Cited by 522 · see citing works
Glucose transporters in adipose tissue, liver, and skeletal muscle in metabolic health and disease
Chadt, A., & Al-Hasani, H. (2020) · Pflügers Archiv - European Journal of Physiology · doi:10.1007/s00424-020-02417-x · Cited by 510 · see citing works
Animal models of non‐alcoholic fatty liver disease: current perspectives and recent advances
Lau, J. K. C., Zhang, X., & Yu, J. (2017) · The Journal of Pathology · doi:10.1002/path.4829 · Cited by 373 · cited 1× · see citing works
“We employed well-established MASLD mouse models to validate our in vitro 3D dynamic MASLD models utilizing 7-week-old male C57BL/6J mice with a high-fat diet feeding [ 16 ].”
Ren, Y., Zhao, H., Yin, C., Lan, X., Wu, L., Du, X., Griffiths, H. R., & Gao, D. (2022) · Frontiers in Endocrinology · doi:10.3389/fendo.2022.873699 · Cited by 260 · see citing works
Nonalcoholic fatty liver disease in adults: current concepts in etiology, outcomes, and management
Marjot, T., Moolla, A., Cobbold, J. F., Hodson, L., & Tomlinson, J. W. (2020) · Endocrine Reviews · doi:10.1210/endrev/bnz009 · Cited by 228 · see citing works
Investigating Nonalcoholic Fatty Liver Disease in a Liver-on-a-Chip Microfluidic Device
Gori, M., Simonelli, M. C., Giannitelli, S. M., Businaro, L., Trombetta, M., & Rainer, A. (2016) · PLoS ONE · doi:10.1371/journal.pone.0159729 · Cited by 177 · see citing works
Effect of intracellular lipid accumulation in a new model of non-alcoholic fatty liver disease
Chávez-Tapia, N. C., Rosso, N., & Tiribelli, C. (2012) · BMC Gastroenterology · doi:10.1186/1471-230x-12-20 · Cited by 138 · see citing works
In Vitro Models for Non-Alcoholic Fatty Liver Disease: Emerging Platforms and Their Applications
Ramos, M. J., Bandiera, L., Menolascina, F., & Fallowfield, J. A. (2022) · iScience · doi:10.1016/j.isci.2021.103549 · Cited by 127 · see citing works
Hepatic transcriptional responses to fasting and feeding
Bideyan, L., Nagari, R., & Tontonoz, P. (2021) · Genes & Development · doi:10.1101/gad.348340.121 · Cited by 126 · see citing works
In Vitro and In Vivo Models of Non-Alcoholic Fatty Liver Disease: A Critical Appraisal
Soret, P.-A., Magusto, J., Housset, C., & Gautheron, J. (2020) · Journal of Clinical Medicine · doi:10.3390/jcm10010036 · Cited by 121 · see citing works
Crosstalk Between Liver Macrophages and Surrounding Cells in Nonalcoholic Steatohepatitis
Li, H., Zhou, Y., Wang, H., Zhang, M., Qiu, P., Zhang, M., Zhang, R., Zhao, Q., & Liu, J. (2020) · Frontiers in Immunology · doi:10.3389/fimmu.2020.01169 · Cited by 114 · see citing works
Human Nonalcoholic Steatohepatitis on a Chip
Freag, M. S., Namgung, B., Reyna Fernandez, M. E., Gherardi, E., Sengupta, S., & Jang, H. L. (2021) · Hepatology Communications · doi:10.1002/hep4.1647 · Cited by 102 · see citing works
Thibaut, R., Gage, M., Pineda‐Torra, I., Chabrier, G., Venteclef, N., & Alzaïd, F. (2021) · FEBS Journal · doi:10.1111/febs.15877 · Cited by 95 · see citing works
Cao, Y., Mai, W., Li, R., Deng, S., Li, L.-J., Zhou, Y.-X., Qin, Q.-S., Zhang, Y., Zhou, X.-G., Han, M., et al. (2022) · Cellular and Molecular Life Sciences · doi:10.1007/s00018-022-04319-w · Cited by 82 · see citing works
Self-citationA Microphysiological System for Studying Nonalcoholic Steatohepatitis
Kostrzewski, T., Maraver, P., Ouro‐Gnao, L., Levi, A., Snow, S., Miedzik, A., Rombouts, K., & Hughes, D. (2020) · Hepatology Communications · doi:10.1002/hep4.1450 · Cited by 77 · see citing works
Breaking the mold: 3D cell cultures reshaping the future of cancer research
Cordeiro, S. Z., Oliveira, B. B. P. P., Valente, R., Ferreira, D., Luz, A., Baptista, P. V., & Fernandes, A. R. (2024) · Frontiers in Cell and Developmental Biology · doi:10.3389/fcell.2024.1507388 · Cited by 64 · see citing works
Cho, H., Kim, H., Lee, K., Lasli, S., Ung, A., Hoffman, T., Nasiri, R., Bandaru, P., Ahadian, S., Dokmeci, M. R., et al. (2021) · Small · doi:10.1002/smll.202007425 · Cited by 62 · see citing works
Ströbel, S., Kostadinova, R. M., Fiaschetti‐Egli, K., Rupp, J., Bieri, M., Pawłowska, A., Busler, D., Hofstetter, T., Sanchez, K., Grepper, S., et al. (2021) · Scientific Reports · doi:10.1038/s41598-021-01951-7 · Cited by 58 · see citing works
Mechanisms of hepatic and renal injury in lipid metabolism disorders in metabolic syndrome
Rong, J., Zhang, Z., Peng, X., Li, P., Zhao, T., & Zhong, Y. (2024) · International Journal of Biological Sciences · doi:10.7150/ijbs.100394 · Cited by 54 · see citing works
Animal Models of Fibrosis in Nonalcoholic Steatohepatitis: Do They Reflect Human Disease?
Ipsen, D. H., Lykkesfeldt, J., & Tveden-Nyborg, P. (2020) · Advances in Nutrition · doi:10.1093/advances/nmaa081 · Cited by 51 · see citing works
Järvinen, E., Hammer, H., Poetz, O., Ingelman‐Sundberg, M., & Stage, T. B. (2023) · Clinical Pharmacology & Therapeutics · doi:10.1002/cpt.2887 · Cited by 51 · see citing works
State-of-the-art liver disease research using liver-on-a-chip
Deguchi, S., & Takayama, K. (2022) · Inflammation and Regeneration · doi:10.1186/s41232-022-00248-0 · Cited by 50 · see citing works
Koui, Y., Himeno, M., Mori, Y., Nakano, Y., Saijou, E., Tanimizu, N., Kamiya, Y., Anzai, H., Maeda, N., Wang, L. L., et al. (2021) · Stem Cell Reports · doi:10.1016/j.stemcr.2021.11.002 · Cited by 46 · see citing works
Zhou, C., Shen, Z., Shen, B., Dai, W., Sun, Z., Guo, Y., Xu, X., Wang, J., Lu, J., Zhang, Q., et al. (2023) · Biochimica et Biophysica Acta (BBA) - Molecular Basis of Disease · doi:10.1016/j.bbadis.2023.166810 · Cited by 45 · see citing works
Zhu, H., Zhao, S., Zhao, S., Yang, S., Jiang, K., Li, S., Kang, Y., Yang, Z., Shen, J., Shen, S., et al. (2024) · Metabolism · doi:10.1016/j.metabol.2024.155914 · Cited by 41 · see citing works
Brandauer, K., Schweinitzer, S., Lorenz, A., Krauß, J., Schobesberger, S., Frauenlob, M., & Ertl, P. (2025) · Lab on a Chip · doi:10.1039/d4lc01011f · Cited by 39 · see citing works
Tanaka, M., Takahashi, S., Higashiura, Y., Sakai, A., Koyama, M., Saitoh, S., Shimamoto, K., Ohnishi, H., & Furuhashi, M. (2021) · Journal of Diabetes Investigation · doi:10.1111/jdi.13735 · Cited by 33 · see citing works
The Role of CTGF in Liver Fibrosis Induced in 3D Human Liver Spheroids
Redenšek, S., van Riet, S., Nordling, Å., & Ingelman‐Sundberg, M. (2023) · Cells · doi:10.3390/cells12020302 · Cited by 33 · see citing works
A Model of Hepatic Steatosis with Declined Viability and Function in a Liver-Organ-on-a-Chip
Wiriyakulsit, N., Keawsomnuk, P., Thongin, S., Ketsawatsomkron, P., & Muta, K. (2023) · Scientific Reports · doi:10.1038/s41598-023-44198-0 · Cited by 29 · see citing works
Developing Liver Microphysiological Systems for Biomedical Applications
Wang, J., Wu, X., Zhao, J., Ren, H., & Zhao, Y. (2024) · Advanced Healthcare Materials · doi:10.1002/adhm.202302217 · Cited by 27 · see citing works
Modelling fatty liver disease with mouse liver-derived multicellular spheroids
van Os, E. A., Cools, L., Eysackers, N., Szafranska, K., Smout, A., Verhulst, S., Reynaert, H., McCourt, P., Mannaerts, I., & van Grunsven, L. A. (2022) · Biomaterials · doi:10.1016/j.biomaterials.2022.121817 · Cited by 25 · see citing works
Holmgren, G., Ulfenborg, B., Asplund, A., Toet, K. H., Andersson, C. X., Hammarstedt, A., Hanemaaijer, R., Küppers-Munther, B., & Synnergren, J. (2020) · International Journal of Molecular Sciences · doi:10.3390/ijms21020469 · Cited by 24 · see citing works
Bogdanos, D. P., Gao, B., & Gershwin, M. E. (2013) · Comprehensive physiology · doi:10.1002/j.2040-4603.2013.tb00503.x · Cited by 24 · see citing works
Insights into Macrophage/Monocyte-Endothelial Cell Crosstalk in the Liver: A Role for Trem-2
Coelho, I., Duarte, N., Macedo, M. P., & Penha‐Gonçalves, C. (2021) · Journal of Clinical Medicine · doi:10.3390/jcm10061248 · Cited by 22 · see citing works
Kermanizadeh, A., Brown, D. M., & Stone, V. (2019) · PLoS ONE · doi:10.1371/journal.pone.0220974 · Cited by 15 · see citing works
Organoids and tissue/organ chips
Seán, G. B., Banes, A. J., & Gangaraju, R. (2024) · Stem Cell Research & Therapy · doi:10.1186/s13287-024-03859-1 · Cited by 9 · see citing works
Karnawat, K., Parthasarathy, R., Sakhrie, M., Karthik, H., Krishna, K. V., & Balachander, G. M. (2024) · Journal of Materials Chemistry B · doi:10.1039/d4tb00738g · Cited by 8 · see citing works
Goswami, Y., Baghel, A., Sharma, G., Sharma, P. K., Biswas, S., Yadav, R., Garg, P. K., Shalimar, S., & Tandon, R. (2024) · Journal of Clinical and Experimental Hepatology · doi:10.1016/j.jceh.2024.102463 · Cited by 6 · see citing works
OCT1 (SLC22A1) transporter kinetics and regulation in primary human hepatocyte 3D spheroids
Mickols, E., Meyer, A., Handin, N., Stüwe, M., Eriksson, J., Rudfeldt, J., Blom, K., Fryknäs, M., Sellin, M. E., Lauschke, V. M., et al. (2024) · Scientific Reports · doi:10.1038/s41598-024-67192-6 · Cited by 5 · see citing works
Integrative Transcriptomics and Proteomics Analysis Reveals THRSP’s Role in Lipid Metabolism
Li, Y., Xu, K., Zhou, A., Xu, Z., Wu, J., Peng, X., Mei, S., & Chen, H. (2024) · Genes · doi:10.3390/genes15121562 · Cited by 5 · see citing works
The Hollow Porous Sphere Cell Carrier for the Dynamic Three-Dimensional Cell Culture
Gao, W., Xiao, L., Wang, J., Mu, Y., Mendhi, J., Gao, W., Li, Z., Yarlagadda, P. K. D. V., Wu, C., & Xiao, Y. (2022) · Tissue Engineering Part C Methods · doi:10.1089/ten.tec.2022.0137 · Cited by 5 · see citing works
Self-citationXing, H., Wu, Z. H., Jiang, K., Yuan, G., Guo, Z., Yu, S., He, S., & Zhong, F. (2024) · Scientific Reports · doi:10.1038/s41598-024-71311-8 · Cited by 3 · see citing works
Flagged first, then by citations
Research works printed but not linked: 3
- [12]Aasadollahei N, Rezaei N, Golroo R, Agarwal T, Vosough M, Piryaei A. Bioengineering liver microtissues for modeling non-alcoholic fatty liver disease. EXCLI J. 2023; 22: 367– 391. 37223084 PMC10201011
- [33]Duriez M, Jacquet A, Hoet L, Roche S, Bock MD, Rocher C, Haussy G, Vigé X, Bocskei Z, Slavnic T, et al. A 3D human liver model of nonalcoholic steatohepatitis. J Clin Transl Hepatol. 2020; 8(4): 359– 370. 33447518 PMC7782122
- [38]Moreno-Vedia J, Girona J, Ibarretxe D, Masana L, Rodríguez-Calvo R. Unveiling the role of the fatty acid binding protein 4 in the metabolic-associated fatty liver disease. Biomedicine. 2022; 10(1): Article 197. PMC8773613 35052876
Cited by
Sorted by citations
The HepG2 Cell Line as a Model for Studying Metabolic Dysfunction-Associated Steatotic Liver Disease
Kotlyarova, A., Iskrina, A., & Kotlyarov, S. (2026) · International journal of molecular sciences · doi:10.3390/ijms27083399 · Cited by 5 · see citing works
Comparison and Characteristics of MASLD Mouse Models
Wei, L., 高春辰, & Qin, H. (2026) · Biomedicines · doi:10.3390/biomedicines14040895 · Cited by 3 · see citing works
Котлярова, А. А., & Kotlyarov, S. N. (2026) · International Journal of Molecular Sciences · doi:10.3390/ijms27104453 · Cited by 1 · see citing works
Zhang, W., Wang, Y., Liu, Y., Zheng, X., Wang, T., Kwan, X., Liu, R., Liu, Q., Zhuang, H., Liang, H., et al. (2026) · Hepatic Medicine Evidence and Research · doi:10.2147/hmer.s605761
Aina, K. O. (2026) · Biochemistry and Biophysics Reports · doi:10.1016/j.bbrep.2026.102616
ATR-FTIR Assessment of the Antioxidant and Anti-Inflammatory Effects of (−)-Epicatechin in MASLD
Hefer, M., Filipović, N., Šarić, S., Malatesti, N., Petrović, A., Kovač-Andrić, E., & Smolić, M. (2026) · ACS Omega · doi:10.1021/acsomega.5c09978
Orbital hybridization-mediated nanozymes reverse cellular senescence for aged bone regeneration
Wang, Y., Cao, J., Wang, X., Liu, T., Jiang, H., Jiang, J., Wang, Y., Yuan, H., Wu, S., & Wu, F. (2026) · Biomaterials · doi:10.1016/j.biomaterials.2026.124352
Cholesterol metabolic rewiring shapes immune remodeling across hepatocarcinogenesis
Ma, W., Yan, F., Cheng, Y., Zhang, G., Mu, Y., Zhang, Z., Liang, X., 甘建新, Yang, W., Zhu, W., et al. (2026) · Frontiers in Immunology · doi:10.3389/fimmu.2026.1884948
Xiu, H., Li, T., Wen, J., Li, Z. B., Wu, J., Fu, X., Zhang, C., Yang, Z., Yao, X., & Xu, Z. (2026) · Clinical Pharmacology & Therapeutics · doi:10.1002/cpt.70416
Vitamin E treatment modulates steatotic and attenuates oxidative stress in a MASLD in vitro model
Teymoorian, S. K., Nouri, K., Choshali, M. A., Hassan, M., Rismani, E., Vosough, M., & Piryaei, A. (2026) · Molecular Biology Reports · doi:10.1007/s11033-026-12691-0
- Authors
- Huang, Z., Li, L., Dudley, K., Xiao, L., Huang, G., Subramaniam, V. N., Chen, C., Crawford, R., & Xiao, Y.
- Affiliations
- Griffith University (AU)
- Queensland University of Technology (AU)
- The University of Queensland (AU)
- Corresponding author
- Xiao, Yin
- ORCID
- Venue
- BME Frontiers
- Published
- 2025-01
- Volume
- 6
- Pages
- 0181-0181
- ISSN
- 2765-8031 (ISSN-L)
- PMC
- PMC12480745
- OpenAlex
- W4413941181
- Cited by
- 10
- References
- 46
- Citations by year
- 2026: 10
- FWCI
- 6.93
- Type
- journal article
- Language
- en
- Open access
- diamond
- doi.org
- Licence
- cc-by
- Locations
- BME Frontiers (publishedVersion, cc-by)
- Europe PMC (PubMed Central) (publishedVersion, cc-by)
- PubMed (publishedVersion)
- PubMed Central (publishedVersion, cc-by)
- DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion, cc-by-sa)
- Griffith Research Online (submittedVersion, cc-by)
- Funders
- National Health and Medical Research Council (APP2000647)
- Topics
- Health Sciences › Medicine › Epidemiology › Liver Disease Diagnosis and Treatment
- Health Sciences › Medicine › Hepatology › Liver physiology and pathology
- Physical Sciences › Engineering › Biomedical Engineering › 3D Printing in Biomedical Research
- Keywords
- non-alcoholic fatty liver disease, MASLD progression, mini-bioreactors, hepatocytes, macrophages, hepatic stellate cells, steatohepatitis, liver fibrosis, hepatocyte lipid metabolism, in vitro disease model
About these checks
Registry, retraction, Cites and statistics results are computed, not judged, with one exception: a printed reference that no index lists is read once by a model before it counts as a citation error. Claims are extracted by a model and kept only when their quote appears verbatim in the text; each cited source is then checked, with a model’s help, for whether it says what the claim relies on. Those verdicts can be wrong — read the claim’s quote and the cited source before relying on one.
Statistics and arithmetic are recomputed, not judged.
Confidence bands are shown only for fields calibrated on our annotated test set. None is calibrated yet, so no band is shown.
What counts as a claim ›How we measure confidence ›How we check this ›
Are you an author, or do you see a problem with this page? Request a review
