Three-Dimensional Liver Culture Systems to Maintain Primary Hepatic Properties for Toxicological Analysis In Vitro
Abstract
:1. Introduction
2. Human Hepatocyte In Vitro Models
- Albumin synthesis, urea production, and glycogen storage
- Expression of phase I enzymes involved in drug metabolism (CYP1A2, CYP2B6, CYP2C9, CYP2C19, CYP2D6, CYP3A4)
- Expression of phase II enzymes such as UDP-glucuronyl transferases (UGTs), glutathione-S-transferases (GSTs) or sulfotransferases (SULTs)
- Cellular polarization marked by the expression of F-actin and transporter molecules (also referred to as phase III) such as MRP2, Pgp, BSEP, OCT1, or OATP1
2.1. Primary Human Hepatocytes
2.2. HepG2 Cells
2.3. HepaRG Cells
2.4. iPSC-Derived Hepatocytes
2.5. Upcyte Hepatocytes/Primary-Like Hepatocytes
2.6. Summary of Human Liver In Vitro Models
3. Three-Dimensional Culture Models for Human Hepatocytes
3.1. Liver Spheroid and Organoid Culture
3.1.1. Spheroids from Primary Human Hepatocytes
3.1.2. Spheroids from HepG2 Cells
3.1.3. Spheroids from HepaRG Cells
3.1.4. Spheroids from iPSC-Derived Hepatocytes
3.1.5. Spheroids from Upcyte and Primary-Like Hepatocytes
3.1.6. Organoids from Human Primary and iPSC-Derived Hepatocytes
3.2. Perfused Bioreactors and Liver-on-a-Chip Models
3.2.1. Perfused Bioreactors with Primary Human Hepatocytes
3.2.2. Perfused Bioreactors with HepG2 Cells
3.2.3. Perfused Bioreactors with HepaRG Cells
3.2.4. Perfused Bioreactors with iPSC-Derived and Upcyte Hepatocytes
4. Conclusions and Outlook
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Conflicts of Interest
References
- Kiamehr, M.; Heiskanen, L.; Laufer, T.; Düsterloh, A.; Kahraman, M.; Käkelä, R.; Laaksonen, R.; Aalto-Setälä, K. Dedifferentiation of Primary Hepatocytes is Accompanied with Reorganization of Lipid Metabolism Indicated by Altered Molecular Lipid and miRNA Profiles. Int. J. Mol. Sci. 2019, 20, 2910. [Google Scholar] [CrossRef] [Green Version]
- Lauschke, V.M.; Vorrink, S.U.; Moro, S.M.L.; Rezayee, F.; Nordling, Å.; Hendriks, D.F.G.; Bell, C.C.; Sison-Young, R.; Park, B.K.; Goldring, C.E.; et al. Massive rearrangements of cellular MicroRNA signatures are key drivers of hepatocyte dedifferentiation. Hepatology 2016, 64, 1743–1756. [Google Scholar] [CrossRef] [Green Version]
- Fisher, K.; Vuppalanchi, R.; Saxena, R. Drug-induced liver injury. Arch. Pathol. Lab. Med. 2015, 139, 876–887. [Google Scholar] [CrossRef]
- Lasser, K.E.; Allen, P.D.; Woolhandler, S.J.; Himmelstein, D.U.; Wolfe, S.M.; Bor, D.H. Timing of new black box warnings and withdrawals for prescription medications. JAMA 2002, 287, 2215–2220. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Russo, M.W.; Galanko, J.A.; Shrestha, R.; Fried, M.W.; Watkins, P. Liver transplantation for acute liver failure from drug induced liver injury in the United States. Liver Transpl. 2004, 10, 1018–1023. [Google Scholar] [CrossRef]
- Ostapowicz, G.; Fontana, R.J.; Schiødt, F.V.; Larson, A.; Davern, T.J.; Han, S.H.; McCashland, T.M.; Shakil, A.O.; Hay, J.E.; Hynan, L.; et al. Results of a prospective study of acute liver failure at 17 tertiary care centers in the United States. Ann. Intern. Med. 2002, 137, 947–954. [Google Scholar] [CrossRef]
- Zhou, Y.; Shen, J.X.; Lauschke, V.M. Comprehensive Evaluation of Organotypic and Microphysiological Liver Models for Prediction of Drug-Induced Liver Injury. Front. Pharmacol. 2019, 10, 1093. [Google Scholar] [CrossRef]
- Chapman, K.L.; Holzgrefe, H.; Black, L.E.; Brown, M.; Chellman, G.; Copeman, C.; Couch, J.; Creton, S.; Gehen, S.; Hoberman, A.; et al. Pharmaceutical toxicology: Designing studies to reduce animal use, while maximizing human translation. Regul. Toxicol. Pharmacol. 2013, 66, 88–103. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Aleksunes, L.M.; Campion, S.N.; Goedken, M.J.; Manautou, J.E. Acquired resistance to acetaminophen hepatotoxicity is associated with induction of multidrug resistance-associated protein 4 (Mrp4) in proliferating hepatocytes. Toxicol. Sci. 2008, 104, 261–273. [Google Scholar] [CrossRef] [PubMed]
- Kozyra, M.; Ingelman-Sundberg, M.; Lauschke, V.M. Rare genetic variants in cellular transporters, metabolic enzymes, and nuclear receptors can be important determinants of interindividual differences in drug response. Genet. Med. 2017, 19, 20–29. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Zhou, Y.; Ingelman-Sundberg, M.; Lauschke, V.M. Worldwide Distribution of Cytochrome P450 Alleles: A Meta-analysis of Population-scale Sequencing Projects. Clin. Pharmacol. Ther. 2017, 102, 688–700. [Google Scholar] [CrossRef] [Green Version]
- López-Terrada, D.; Cheung, S.W.; Finegold, M.J.; Knowles, B.B. Hep G2 is a hepatoblastoma-derived cell line. Hum. Pathol. 2009, 40, 1512–1515. [Google Scholar] [CrossRef] [PubMed]
- Aden, D.P.; Fogel, A.; Plotkin, S.; Damjanov, I.; Knowles, B.B. Controlled synthesis of HBsAg in a differentiated human liver carcinoma-derived cell line. Nature 1979, 282, 615–616. [Google Scholar] [CrossRef]
- Busso, N.; Chesne, C.; Delers, F.; Morel, F.; Guillouzo, A. Transforming growth-factor-beta (TGF-beta) inhibits albumin synthesis in normal human hepatocytes and in hepatoma HepG2 cells. Biochem. Biophys. Res. Commun. 1990, 171, 647–654. [Google Scholar] [CrossRef]
- Meier, M.; Klein, H.H.; Kramer, J.; Drenckhan, M.; Schütt, M. Calpain inhibition impairs glycogen syntheses in HepG2 hepatoma cells without altering insulin signaling. J. Endocrinol. 2007, 193, 45–51. [Google Scholar] [CrossRef] [Green Version]
- Gerets, H.H.; Tilmant, K.; Gerin, B.; Chanteux, H.; Depelchin, B.O.; Dhalluin, S.; Atienzar, F.A. Characterization of primary human hepatocytes, HepG2 cells, and HepaRG cells at the mRNA level and CYP activity in response to inducers and their predictivity for the detection of human hepatotoxins. Cell Biol. Toxicol. 2012, 28, 69–87. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Westerink, W.M.; Schoonen, W.G. Cytochrome P450 enzyme levels in HepG2 cells and cryopreserved primary human hepatocytes and their induction in HepG2 cells. Toxicol. In Vitro 2007, 21, 1581–1591. [Google Scholar] [CrossRef]
- Steinbrecht, S.; Pfeifer, N.; Herzog, N.; Katzenberger, N.; Schulz, C.; Kammerer, S.; Küpper, J.H. HepG2-1A2 C2 and C7: Lentivirus vector-mediated stable and functional overexpression of cytochrome P450 1A2 in human hepatoblastoma cells. Toxicol. Lett. 2020, 319, 155–159. [Google Scholar] [CrossRef] [PubMed]
- Steinbrecht, S.; König, R.; Schmidtke, K.U.; Herzog, N.; Scheibner, K.; Krüger-Genge, A.; Jung, F.; Kammerer, S.; Küpper, J.H. Metabolic activity testing can underestimate acute drug cytotoxicity as revealed by HepG2 cell clones overexpressing cytochrome P450 2C19 and 3A4. Toxicology 2019, 412, 37–47. [Google Scholar] [CrossRef] [PubMed]
- Xuan, J.; Chen, S.; Ning, B.; Tolleson, W.H.; Guo, L. Development of HepG2-derived cells expressing cytochrome P450s for assessing metabolism-associated drug-induced liver toxicity. Chem. Biol. Interact. 2016, 255, 63–73. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Herzog, N.; Katzenberger, N.; Martin, F.; Schmidtke, K.-U.; Küpper, J.-H. Generation of cytochrome P450 3A4-overexpressing HepG2 cell clones for standardization of hepatocellular testosterone 6β-hydroxylation activity. J. Cell. Biotechnol. 2015, 1, 15–26. [Google Scholar] [CrossRef] [Green Version]
- Yoshitomi, S.; Ikemoto, K.; Takahashi, J.; Miki, H.; Namba, M.; Asahi, S. Establishment of the transformants expressing human cytochrome P450 subtypes in HepG2, and their applications on drug metabolism and toxicology. Toxicol. In Vitro 2001, 15, 245–256. [Google Scholar] [CrossRef]
- Gripon, P.; Rumin, S.; Urban, S.; Le Seyec, J.; Glaise, D.; Cannie, I.; Guyomard, C.; Lucas, J.; Trepo, C.; Guguen-Guillouzo, C. Infection of a human hepatoma cell line by hepatitis B virus. Proc. Natl. Acad. Sci. USA 2002, 99, 15655–15660. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Jennen, D.G.; Magkoufopoulou, C.; Ketelslegers, H.B.; van Herwijnen, M.H.; Kleinjans, J.C.; van Delft, J.H. Comparison of HepG2 and HepaRG by whole-genome gene expression analysis for the purpose of chemical hazard identification. Toxicol. Sci. 2010, 115, 66–79. [Google Scholar] [CrossRef] [Green Version]
- Aninat, C.; Piton, A.; Glaise, D.; Le Charpentier, T.; Langouët, 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. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Klein, S.; Mueller, D.; Schevchenko, V.; Noor, F. Long-term maintenance of HepaRG cells in serum-free conditions and application in a repeated dose study. J. Appl. Toxicol. 2014, 34, 1078–1086. [Google Scholar] [CrossRef]
- Jackson, J.P.; Li, L.; Chamberlain, E.D.; Wang, H.; Ferguson, S.S. Contextualizing Hepatocyte Functionality of Cryopreserved HepaRG Cell Cultures. Drug Metab. Dispos. 2016, 44, 1463–1479. [Google Scholar] [CrossRef] [Green Version]
- Guillouzo, A.; Corlu, A.; Aninat, C.; Glaise, D.; Morel, F.; Guguen-Guillouzo, C. The human hepatoma HepaRG cells: A highly differentiated model for studies of liver metabolism and toxicity of xenobiotics. Chem. Biol. Interact. 2007, 168, 66–73. [Google Scholar] [CrossRef] [PubMed]
- Zanger, U.M.; Schwab, M. Cytochrome P450 enzymes in drug metabolism: Regulation of gene expression, enzyme activities, and impact of genetic variation. Pharmacol. Ther. 2013, 138, 103–141. [Google Scholar] [CrossRef]
- Goldring, C.; Antoine, D.J.; Bonner, F.; Crozier, J.; Denning, C.; Fontana, R.J.; Hanley, N.A.; Hay, D.C.; Ingelman-Sundberg, M.; Juhila, S. Stem cell–derived models to improve mechanistic understanding and prediction of human drug-induced liver injury. Hepatology 2017, 65, 710–721. [Google Scholar] [CrossRef]
- Kia, R.; Sison, R.L.; Heslop, J.; Kitteringham, N.R.; Hanley, N.; Mills, J.S.; Park, B.K.; Goldring, C.E. Stem cell-derived hepatocytes as a predictive model for drug-induced liver injury: Are we there yet? Br. J. Clin. Pharmacol. 2013, 75, 885–896. [Google Scholar] [CrossRef] [Green Version]
- Freyer, N.; Knöspel, F.; Strahl, N.; Amini, L.; Schrade, P.; Bachmann, S.; Damm, G.; Seehofer, D.; Jacobs, F.; Monshouwer, M.; et al. Hepatic Differentiation of Human Induced Pluripotent Stem Cells in a Perfused Three-Dimensional Multicompartment Bioreactor. Biores. Open Access 2016, 5, 235–248. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Kang, S.J.; Lee, H.M.; Park, Y.I.; Yi, H.; Lee, H.; So, B.; Song, J.Y.; Kang, H.G. Chemically induced hepatotoxicity in human stem cell-induced hepatocytes compared with primary hepatocytes and HepG2. Cell Biol. Toxicol. 2016, 32, 403–417. [Google Scholar] [CrossRef]
- Baxter, M.; Withey, S.; Harrison, S.; Segeritz, C.-P.; Zhang, F.; Atkinson-Dell, R.; Rowe, C.; Gerrard, D.T.; Sison-Young, R.; Jenkins, R. Phenotypic and functional analyses show stem cell-derived hepatocyte-like cells better mimic fetal rather than adult hepatocytes. J. Hepatol. 2015, 62, 581–589. [Google Scholar] [CrossRef]
- Schwartz, R.E.; Fleming, H.E.; Khetani, S.R.; Bhatia, S.N. Pluripotent stem cell-derived hepatocyte-like cells. Biotechnol. Adv. 2014, 32, 504–513. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Takayama, K.; Morisaki, Y.; Kuno, S.; Nagamoto, Y.; Harada, K.; Furukawa, N.; Ohtaka, M.; Nishimura, K.; Imagawa, K.; Sakurai, F.; et al. Prediction of interindividual differences in hepatic functions and drug sensitivity by using human iPS-derived hepatocytes. Proc. Natl. Acad. Sci. USA 2014, 111, 16772–16777. [Google Scholar] [CrossRef] [Green Version]
- Ulvestad, M.; Nordell, P.; Asplund, A.; Rehnström, M.; Jacobsson, S.; Holmgren, G.; Davidson, L.; Brolén, G.; Edsbagge, J.; Björquist, P.; et al. Drug metabolizing enzyme and transporter protein profiles of hepatocytes derived from human embryonic and induced pluripotent stem cells. Biochem. Pharmacol. 2013, 86, 691–702. [Google Scholar] [CrossRef] [PubMed]
- Gao, X.; Liu, Y. A transcriptomic study suggesting human iPSC-derived hepatocytes potentially offer a better in vitro model of hepatotoxicity than most hepatoma cell lines. Cell Biol. Toxicol. 2017, 33, 407–421. [Google Scholar] [CrossRef] [PubMed]
- Holmgren, G.; Ulfenborg, B.; Asplund, A.; Toet, K.; Andersson, C.X.; Hammarstedt, A.; Hanemaaijer, R.; Küppers-Munther, B.; Synnergren, J. Characterization of Human Induced Pluripotent Stem Cell-Derived Hepatocytes with Mature Features and Potential for Modeling Metabolic Diseases. Int. J. Mol. Sci. 2020, 21, 469. [Google Scholar] [CrossRef] [Green Version]
- Kammerer, S.; Küpper, J.H. Optimized protocol for induction of cytochrome P450 enzymes 1A2 and 3A4 in human primary-like hepatocyte cell strain HepaFH3 to study in vitro toxicology. Clin. Hemorheol. Microcirc. 2018, 70, 563–571. [Google Scholar] [CrossRef]
- Herzog, N.; Hansen, M.; Miethbauer, S.; Schmidtke, K.U.; Anderer, U.; Lupp, A.; Sperling, S.; Seehofer, D.; Damm, G.; Scheibner, K. Primary-like human hepatocytes genetically engineered to obtain proliferation competence display hepatic differentiation characteristics in monolayer and organotypical spheroid cultures. Cell Biol. Int. 2016, 40, 341–353. [Google Scholar] [CrossRef]
- Nörenberg, A.; Heinz, S.; Scheller, K.; Hewitt, N.J.; Braspenning, J.; Ott, M. Optimization of upcyte® human hepatocytes for the in vitro micronucleus assay. Mutat. Res. Genet. Toxicol. Environ. Mutagenesis 2013, 758, 69–79. [Google Scholar] [CrossRef]
- Burkard, A.; Dähn, C.; Heinz, S.; Zutavern, A.; Sonntag-Buck, V.; Maltman, D.; Przyborski, S.; Hewitt, N.J.; Braspenning, J. Generation of proliferating human hepatocytes using upcyte® technology: Characterisation and applications in induction and cytotoxicity assays. Xenobiotica 2012, 42, 939–956. [Google Scholar] [CrossRef] [PubMed]
- Levy, G.; Bomze, D.; Heinz, S.; Ramachandran, S.D.; Noerenberg, A.; Cohen, M.; Shibolet, O.; Sklan, E.; Braspenning, J.; Nahmias, Y. Long-term culture and expansion of primary human hepatocytes. Nat. Biotechnol. 2015, 33, 1264–1271. [Google Scholar] [CrossRef] [PubMed]
- Ramachandran, S.D.; Vivarès, A.; Klieber, S.; Hewitt, N.J.; Muenst, B.; Heinz, S.; Walles, H.; Braspenning, J. Applicability of second-generation upcyte® human hepatocytes for use in CYP inhibition and induction studies. Pharmacol. Res. Perspect. 2015, 3, e00161. [Google Scholar] [CrossRef]
- Tolosa, L.; Gómez-Lechón, M.J.; López, S.; Guzmán, C.; Castell, J.V.; Donato, M.T.; Jover, R. Human Upcyte Hepatocytes: Characterization of the Hepatic Phenotype and Evaluation for Acute and Long-Term Hepatotoxicity Routine Testing. Toxicol. Sci. 2016, 152, 214–229. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Fu, G.B.; Huang, W.J.; Zeng, M.; Zhou, X.; Wu, H.P.; Liu, C.C.; Wu, H.; Weng, J.; Zhang, H.D.; Cai, Y.C.; et al. Expansion and differentiation of human hepatocyte-derived liver progenitor-like cells and their use for the study of hepatotropic pathogens. Cell Res. 2019, 29, 8–22. [Google Scholar] [CrossRef] [PubMed]
- Qiao, S.; Feng, S.; Wu, Z.; He, T.; Ma, C.; Peng, Z.; Tian, E.; Pan, G. Functional Proliferating Human Hepatocytes: In Vitro Hepatocyte Model for Drug Metabolism, Excretion, and Toxicity. Drug Metab. Dispos. 2021, 49, 305–313. [Google Scholar] [CrossRef] [PubMed]
- Vinken, M.; Papeleu, P.; Snykers, S.; De Rop, E.; Henkens, T.; Chipman, J.K.; Rogiers, V.; Vanhaecke, T. Involvement of cell junctions in hepatocyte culture functionality. Crit. Rev. Toxicol. 2006, 36, 299–318. [Google Scholar] [CrossRef]
- Riede, J.; Wollmann, B.M.; Molden, E.; Ingelman-Sundberg, M. Primary human hepatocyte spheroids as an in vitro tool for investigating drug compounds with low clearance. Drug Metab. Dispos. 2021. [Google Scholar] [CrossRef]
- Rose, S.; Ezan, F.; Cuvellier, M.; Bruyère, A.; Legagneux, V.; Langouët, S.; Baffet, G. Generation of proliferating human adult hepatocytes using optimized 3D culture conditions. Sci. Rep. 2021, 11, 515. [Google Scholar] [CrossRef]
- Mizoi, K.; Hosono, M.; Kojima, H.; Ogihara, T. Establishment of a primary human hepatocyte spheroid system for evaluating metabolic toxicity using dacarbazine under conditions of CYP1A2 induction. Drug Metab. Pharmacokinet. 2020, 35, 201–206. [Google Scholar] [CrossRef] [PubMed]
- Bell, C.C.; Hendriks, D.F.G.; Moro, S.M.L.; Ellis, E.; Walsh, J.; Renblom, A.; Fredriksson Puigvert, L.; Dankers, A.C.A.; Jacobs, F.; Snoeys, J.; et al. Characterization of primary human hepatocyte spheroids as a model system for drug-induced liver injury, liver function and disease. Sci. Rep. 2016, 6, 25187. [Google Scholar] [CrossRef] [Green Version]
- Messner, S.; Agarkova, I.; Moritz, W.; Kelm, J.M. Multi-cell type human liver microtissues for hepatotoxicity testing. Arch. Toxicol. 2013, 87, 209–213. [Google Scholar] [CrossRef] [Green Version]
- Kanebratt, K.P.; Janefeldt, A.; Vilén, L.; Vildhede, A.; Samuelsson, K.; Milton, L.; Björkbom, A.; Persson, M.; Leandersson, C.; Andersson, T.B.; et al. Primary Human Hepatocyte Spheroid Model as a 3D In Vitro Platform for Metabolism Studies. J. Pharm. Sci. 2021, 110, 422–431. [Google Scholar] [CrossRef]
- Vorrink, S.U.; Ullah, S.; Schmidt, S.; Nandania, J.; Velagapudi, V.; Beck, O.; Ingelman-Sundberg, M.; Lauschke, V.M. Endogenous and xenobiotic metabolic stability of primary human hepatocytes in long-term 3D spheroid cultures revealed by a combination of targeted and untargeted metabolomics. FASEB J. 2017, 31, 2696–2708. [Google Scholar] [CrossRef] [Green Version]
- Berger, B.; Donzelli, M.; Maseneni, S.; Boess, F.; Roth, A.; Krähenbühl, S.; Haschke, M. Comparison Of Liver Cell Models Using The Basel Phenotyping Cocktail. Front. Pharmacol. 2016, 7, 443. [Google Scholar] [CrossRef] [Green Version]
- Foster, A.J.; Chouhan, B.; Regan, S.L.; Rollison, H.; Amberntsson, S.; Andersson, L.C.; Srivastava, A.; Darnell, M.; Cairns, J.; Lazic, S.E.; et al. Integrated in vitro models for hepatic safety and metabolism: Evaluation of a human Liver-Chip and liver spheroid. Arch. Toxicol. 2019, 93, 1021–1037. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Bell, C.C.; Lauschke, V.M.; Vorrink, S.U.; Palmgren, H.; Duffin, R.; Andersson, T.B.; Ingelman-Sundberg, M. Transcriptional, Functional, and Mechanistic Comparisons of Stem Cell–Derived Hepatocytes, HepaRG Cells, and Three-Dimensional Human Hepatocyte Spheroids as Predictive In Vitro Systems for Drug-Induced Liver Injury. Drug Metab. Dispos. 2017, 45, 419–429. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Bell, C.C.; Dankers, A.C.A.; Lauschke, V.M.; Sison-Young, R.; Jenkins, R.; Rowe, C.; Goldring, C.E.; Park, K.; Regan, S.L.; Walker, T.; et al. Comparison of Hepatic 2D Sandwich Cultures and 3D Spheroids for Long-term Toxicity Applications: A Multicenter Study. Toxicol. Sci. 2018, 162, 655–666. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Vorrink, S.U.; Zhou, Y.; Ingelman-Sundberg, M.; Lauschke, V.M. Prediction of Drug-Induced Hepatotoxicity Using Long-Term Stable Primary Hepatic 3D Spheroid Cultures in Chemically Defined Conditions. Toxicol. Sci. Off. J. Soc. Toxicol. 2018, 163, 655–665. [Google Scholar] [CrossRef] [PubMed]
- Hendriks, D.F.; Fredriksson Puigvert, L.; Messner, S.; Mortiz, W.; Ingelman-Sundberg, M. Hepatic 3D spheroid models for the detection and study of compounds with cholestatic liability. Sci. Rep. 2016, 6, 35434. [Google Scholar] [CrossRef] [PubMed]
- Kozyra, M.; Johansson, I.; Nordling, Å.; Ullah, S.; Lauschke, V.M.; Ingelman-Sundberg, M. Human hepatic 3D spheroids as a model for steatosis and insulin resistance. Sci. Rep. 2018, 8, 14297. [Google Scholar] [CrossRef] [Green Version]
- Rubiano, A.; Indapurkar, A.; Yokosawa, R.; Miedzik, A.; Rosenzweig, B.; Arefin, A.; Moulin, C.M.; Dame, K.; Hartman, N.; Volpe, D.A.; et al. Characterizing the reproducibility in using a liver microphysiological system for assaying drug toxicity, metabolism, and accumulation. Clin. Transl. Sci. 2021, 14, 1049–1061. [Google Scholar] [CrossRef] [PubMed]
- Kukla, D.A.; Crampton, A.L.; Wood, D.K.; Khetani, S.R. Microscale Collagen and Fibroblast Interactions Enhance Primary Human Hepatocyte Functions in Three-Dimensional Models. Gene Expr. 2020, 20, 1–18. [Google Scholar] [CrossRef] [PubMed]
- Messner, S.; Fredriksson, L.; Lauschke, V.M.; Roessger, K.; Escher, C.; Bober, M.; Kelm, J.M.; Ingelman-Sundberg, M.; Moritz, W. Transcriptomic, Proteomic, and Functional Long-Term Characterization of Multicellular Three-Dimensional Human Liver Microtissues. Appl. In Vitro Toxicol. 2018, 4, 1–12. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Baze, A.; Parmentier, C.; Hendriks, D.F.; Hurrell, T.; Heyd, B.; Bachellier, P.; Schuster, C.; Ingelman-Sundberg, M.; Richert, L. Three-dimensional spheroid primary human hepatocytes in monoculture and coculture with nonparenchymal cells. Tissue Eng. Part C Methods 2018, 24, 534–545. [Google Scholar] [CrossRef]
- Bell, C.C.; Chouhan, B.; Andersson, L.C.; Andersson, H.; Dear, J.W.; Williams, D.P.; Söderberg, M. Functionality of primary hepatic non-parenchymal cells in a 3D spheroid model and contribution to acetaminophen hepatotoxicity. Arch. Toxicol. 2020, 94, 1251–1263. [Google Scholar] [CrossRef] [Green Version]
- Li, F.; Cao, L.; Parikh, S.; Zuo, R. Three-dimensional spheroids with primary human liver cells and differential roles of kupffer cells in drug-induced liver injury. J. Pharm. Sci. 2020, 109, 1912–1923. [Google Scholar] [CrossRef]
- Proctor, W.R.; Foster, A.J.; Vogt, J.; Summers, C.; Middleton, B.; Pilling, M.A.; Shienson, D.; Kijanska, M.; Ströbel, S.; Kelm, J.M.; et al. Utility of spherical human liver microtissues for prediction of clinical drug-induced liver injury. Arch. Toxicol. 2017, 91, 2849–2863. [Google Scholar] [CrossRef]
- Nguyen, D.G.; Funk, J.; Robbins, J.B.; Crogan-Grundy, C.; Presnell, S.C.; Singer, T.; Roth, A.B. Bioprinted 3D primary liver tissues allow assessment of organ-level response to clinical drug induced toxicity in vitro. PLoS ONE 2016, 11, e0158674. [Google Scholar] [CrossRef]
- Tostões, R.M.; Leite, S.B.; Serra, M.; Jensen, J.; Björquist, P.; Carrondo, M.J.T.; Brito, C.; Alves, P.M. Human liver cell spheroids in extended perfusion bioreactor culture for repeated-dose drug testing. Hepatology 2012, 55, 1227–1236. [Google Scholar] [CrossRef] [PubMed]
- Rebelo, S.P.; Costa, R.; Silva, M.M.; Marcelino, P.; Brito, C.; Alves, P.M. Three-dimensional co-culture of human hepatocytes and mesenchymal stem cells: Improved functionality in long-term bioreactor cultures. J. Tissue Eng. Regen. Med. 2017, 11, 2034–2045. [Google Scholar] [CrossRef] [PubMed]
- Štampar, M.; Breznik, B.; Filipič, M.; Žegura, B. Characterization of In Vitro 3D Cell Model Developed from Human Hepatocellular Carcinoma (HepG2) Cell Line. Cells 2020, 9, 2557. [Google Scholar] [CrossRef] [PubMed]
- Štampar, M.; Tomc, J.; Filipič, M.; Žegura, B. Development of in vitro 3D cell model from hepatocellular carcinoma (HepG2) cell line and its application for genotoxicity testing. Arch. Toxicol. 2019, 93, 3321–3333. [Google Scholar] [CrossRef]
- Gaskell, H.; Sharma, P.; Colley, H.E.; Murdoch, C.; Williams, D.P.; Webb, S.D. Characterization of a functional C3A liver spheroid model. Toxicol. Res. 2016, 5, 1053–1065. [Google Scholar] [CrossRef] [Green Version]
- Takahashi, Y.; Hori, Y.; Yamamoto, T.; Urashima, T.; Ohara, Y.; Tanaka, H. 3D spheroid cultures improve the metabolic gene expression profiles of HepaRG cells. Biosci. Rep. 2015, 35. [Google Scholar] [CrossRef]
- Chang, T.T.; Hughes-Fulford, M. Monolayer and spheroid culture of human liver hepatocellular carcinoma cell line cells demonstrate distinct global gene expression patterns and functional phenotypes. Tissue Eng. Part A 2009, 15, 559–567. [Google Scholar] [CrossRef]
- Elje, E.; Hesler, M.; Rundén-Pran, E.; Mann, P.; Mariussen, E.; Wagner, S.; Dusinska, M.; Kohl, Y. The comet assay applied to HepG2 liver spheroids. Mutat. Res./Genet. Toxicol. Environ. Mutagenesis 2019, 845, 403033. [Google Scholar] [CrossRef]
- Štampar, M.; Sedighi Frandsen, H.; Rogowska-Wrzesinska, A.; Wrzesinski, K.; Filipič, M.; Žegura, B. Hepatocellular carcinoma (HepG2/C3A) cell-based 3D model for genotoxicity testing of chemicals. Sci. Total Environ. 2021, 755, 143255. [Google Scholar] [CrossRef]
- Sasaki, K.; Akagi, T.; Asaoka, T.; Eguchi, H.; Fukuda, Y.; Iwagami, Y.; Yamada, D.; Noda, T.; Wada, H.; Gotoh, K. Construction of three-dimensional vascularized functional human liver tissue using a layer-by-layer cell coating technique. Biomaterials 2017, 133, 263–274. [Google Scholar] [CrossRef]
- Mori, N.; Kida, Y.S. Expression of genes involved in drug metabolism differs between perfusable 3D liver tissue and conventional 2D-cultured hepatocellular carcinoma cells. FEBS Open Bio 2020, 10, 1985–2002. [Google Scholar] [CrossRef]
- Kang, H.K.; Sarsenova, M.; Kim, D.H.; Kim, M.S.; Lee, J.Y.; Sung, E.A.; Kook, M.G.; Kim, N.G.; Choi, S.W.; Ogay, V.; et al. Establishing a 3D In Vitro Hepatic Model Mimicking Physiologically Relevant to In Vivo State. Cells 2021, 10, 1268. [Google Scholar] [CrossRef] [PubMed]
- Gori, M.; Giannitelli, S.M.; Torre, M.; Mozetic, P.; Abbruzzese, F.; Trombetta, M.; Traversa, E.; Moroni, L.; Rainer, A. Biofabrication of Hepatic Constructs by 3D Bioprinting of a Cell-Laden Thermogel: An Effective Tool to Assess Drug-Induced Hepatotoxic Response. Adv. Healthc. Mater. 2020, 9, e2001163. [Google Scholar] [CrossRef]
- Taymour, R.; Kilian, D.; Ahlfeld, T.; Gelinsky, M.; Lode, A. 3D bioprinting of hepatocytes: Core-shell structured co-cultures with fibroblasts for enhanced functionality. Sci. Rep. 2021, 11, 5130. [Google Scholar] [CrossRef] [PubMed]
- Mueller, D.; Krämer, L.; Hoffmann, E.; Klein, S.; Noor, F. 3D organotypic HepaRG cultures as in vitro model for acute and repeated dose toxicity studies. Toxicology In Vitro 2014, 28, 104–112. [Google Scholar] [CrossRef] [PubMed]
- Cuvellier, M.; Ezan, F.; Oliveira, H.; Rose, S.; Fricain, J.C.; Langouët, S.; Legagneux, V.; Baffet, G. 3D culture of HepaRG cells in GelMa and its application to bioprinting of a multicellular hepatic model. Biomaterials 2021, 269, 120611. [Google Scholar] [CrossRef] [PubMed]
- Ott, L.M.; Ramachandran, K.; Stehno-Bittel, L. An Automated Multiplexed Hepatotoxicity and CYP Induction Assay Using HepaRG Cells in 2D and 3D. SLAS DISCOVERY Adv. Sci. Drug Discov. 2017, 22, 614–625. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Wang, Z.; Luo, X.; Anene-Nzelu, C.; Yu, Y.; Hong, X.; Singh, N.H.; Xia, L.; Liu, S.; Yu, H. HepaRG culture in tethered spheroids as an in vitro three-dimensional model for drug safety screening. J. Appl. Toxicol. 2015, 35, 909–917. [Google Scholar] [CrossRef]
- Gunness, P.; Mueller, D.; Shevchenko, V.; Heinzle, E.; Ingelman-Sundberg, M.; Noor, F. 3D Organotypic Cultures of Human HepaRG Cells: A Tool for In Vitro Toxicity Studies. Toxicol. Sci. 2013, 133, 67–78. [Google Scholar] [CrossRef] [PubMed]
- 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.; et al. Three-Dimensional HepaRG Model As An Attractive Tool for Toxicity Testing. Toxicol. Sci. 2012, 130, 106–116. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Nelson, L.J.; Morgan, K.; Treskes, P.; Samuel, K.; Henderson, C.J.; LeBled, C.; Homer, N.; Grant, M.H.; Hayes, P.C.; Plevris, J.N. Human Hepatic HepaRG Cells Maintain an Organotypic Phenotype with High Intrinsic CYP450 Activity/Metabolism and Significantly Outperform Standard HepG2/C3A Cells for Pharmaceutical and Therapeutic Applications. Basic Clin. Pharmacol. Toxicol. 2017, 120, 30–37. [Google Scholar] [CrossRef]
- Mandon, M.; Huet, S.; Dubreil, E.; Fessard, V.; Le Hégarat, L. Three-dimensional HepaRG spheroids as a liver model to study human genotoxicity in vitro with the single cell gel electrophoresis assay. Sci. Rep. 2019, 9, 1–9. [Google Scholar]
- Zhang, C.; Zhang, Q.; Li, J.; Yu, L.; Li, F.; Li, W.; Li, Y.; Peng, H.; Zhao, J.; Carmichael, P.L.; et al. Integration of in vitro data from three dimensionally cultured HepaRG cells and physiologically based pharmacokinetic modeling for assessment of acetaminophen hepatotoxicity. Regul. Toxicol. Pharmacol. 2020, 114, 104661. [Google Scholar] [CrossRef] [PubMed]
- Basharat, A.; Rollison, H.E.; Williams, D.P.; Ivanov, D.P. HepG2 (C3A) spheroids show higher sensitivity compared to HepaRG spheroids for drug-induced liver injury (DILI). Toxicol. Appl. Pharmacol. 2020, 408, 115279. [Google Scholar] [CrossRef] [PubMed]
- Weltin, A.; Hammer, S.; Noor, F.; Kaminski, Y.; Kieninger, J.; Urban, G.A. Accessing 3D microtissue metabolism: Lactate and oxygen monitoring in hepatocyte spheroids. Biosens. Bioelectron. 2017, 87, 941–948. [Google Scholar] [CrossRef] [PubMed]
- Leite, S.B.; Roosens, T.; El Taghdouini, A.; Mannaerts, I.; Smout, A.J.; Najimi, M.; Sokal, E.; Noor, F.; Chesne, C.; van Grunsven, L.A. Novel human hepatic organoid model enables testing of drug-induced liver fibrosis in vitro. Biomaterials 2016, 78, 1–10. [Google Scholar] [CrossRef] [PubMed]
- Rashidi, H.; Luu, N.-T.; Alwahsh, S.M.; Ginai, M.; Alhaque, S.; Dong, H.; Tomaz, R.A.; Vernay, B.; Vigneswara, V.; Hallett, J.M. 3D human liver tissue from pluripotent stem cells displays stable phenotype in vitro and supports compromised liver function in vivo. Arch. Toxicol. 2018, 92, 3117–3129. [Google Scholar] [CrossRef] [Green Version]
- Meier, F.; Freyer, N.; Brzeszczynska, J.; Knöspel, F.; Armstrong, L.; Lako, M.; Greuel, S.; Damm, G.; Ludwig-Schwellinger, E.; Deschl, U.; et al. Hepatic differentiation of human iPSCs in different 3D models: A comparative study. Int. J. Mol. Med. 2017, 40, 1759–1771. [Google Scholar] [CrossRef]
- Takayama, K.; Kawabata, K.; Nagamoto, Y.; Kishimoto, K.; Tashiro, K.; Sakurai, F.; Tachibana, M.; Kanda, K.; Hayakawa, T.; Furue, M.K.; et al. 3D spheroid culture of hESC/hiPSC-derived hepatocyte-like cells for drug toxicity testing. Biomaterials 2013, 34, 1781–1789. [Google Scholar] [CrossRef]
- Lee, G.; Kim, H.; Park, J.Y.; Kim, G.; Han, J.; Chung, S.; Yang, J.H.; Jeon, J.S.; Woo, D.H.; Han, C.; et al. Generation of uniform liver spheroids from human pluripotent stem cells for imaging-based drug toxicity analysis. Biomaterials 2021, 269, 120529. [Google Scholar] [CrossRef]
- Qosa, H.; Ribeiro, A.J.S.; Hartman, N.R.; Volpe, D.A. Characterization of a commercially available line of iPSC hepatocytes as models of hepatocyte function and toxicity for regulatory purposes. J. Pharmacol. Toxicol. Methods 2021, 110, 107083. [Google Scholar] [CrossRef]
- Goulart, E.; de Caires-Junior, L.C.; Telles-Silva, K.A.; Araujo, B.H.S.; Rocco, S.A.; Sforca, M.; de Sousa, I.L.; Kobayashi, G.S.; Musso, C.M.; Assoni, A.F.; et al. 3D bioprinting of liver spheroids derived from human induced pluripotent stem cells sustain liver function and viability in vitro. Biofabrication 2019, 12, 015010. [Google Scholar] [CrossRef]
- Ardalani, H.; Sengupta, S.; Harms, V.; Vickerman, V.; Thomson, J.A.; Murphy, W.L. 3-D culture and endothelial cells improve maturity of human pluripotent stem cell-derived hepatocytes. Acta Biomater. 2019, 95, 371–381. [Google Scholar] [CrossRef]
- Sirenko, O.; Hancock, M.K.; Hesley, J.; Hong, D.; Cohen, A.; Gentry, J.; Carlson, C.B.; Mann, D.A. Phenotypic characterization of toxic compound effects on liver spheroids derived from iPSC using confocal imaging and three-dimensional image analysis. Assay Drug Dev. Technol. 2016, 14, 381–394. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Wang, Z.; Li, W.; Jing, H.; Ding, M.; Fu, G.; Yuan, T.; Huang, W.; Dai, M.; Tang, D.; Zeng, M. Generation of hepatic spheroids using human hepatocyte-derived liver progenitor-like cells for hepatotoxicity screening. Theranostics 2019, 9, 6690. [Google Scholar] [CrossRef] [PubMed]
- Thompson, W.L.; Takebe, T. Generation of multi-cellular human liver organoids from pluripotent stem cells. Methods Cell Biol. 2020, 159, 47–68. [Google Scholar] [CrossRef] [PubMed]
- Hu, H.; Gehart, H.; Artegiani, B.; LÖpez-Iglesias, C.; Dekkers, F.; Basak, O.; van Es, J.; Chuva de Sousa Lopes, S.M.; Begthel, H.; Korving, J.; et al. Long-Term Expansion of Functional Mouse and Human Hepatocytes as 3D Organoids. Cell 2018, 175, 1591–1606.e19. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Huch, M.; Gehart, H.; Van Boxtel, R.; Hamer, K.; Blokzijl, F.; Verstegen, M.M.; Ellis, E.; Van Wenum, M.; Fuchs, S.A.; de Ligt, J. Long-term culture of genome-stable bipotent stem cells from adult human liver. Cell 2015, 160, 299–312. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Mun, S.J.; Hong, Y.H.; Ahn, H.S.; Ryu, J.S.; Chung, K.S.; Son, M.J. Long-Term Expansion of Functional Human Pluripotent Stem Cell-Derived Hepatic Organoids. Int. J. Stem Cells 2020, 13, 279–286. [Google Scholar] [CrossRef] [Green Version]
- Ramli, M.N.B.; Lim, Y.S.; Koe, C.T.; Demircioglu, D.; Tng, W.; Gonzales, K.A.U.; Tan, C.P.; Szczerbinska, I.; Liang, H.; Soe, E.L. Human pluripotent stem cell-derived organoids as models of liver disease. Gastroenterology 2020, 159, 1471–1486.e12. [Google Scholar] [CrossRef]
- Pettinato, G.; Lehoux, S.; Ramanathan, R.; Salem, M.M.; He, L.X.; Muse, O.; Flaumenhaft, R.; Thompson, M.T.; Rouse, E.A.; Cummings, R.D.; et al. Generation of fully functional hepatocyte-like organoids from human induced pluripotent stem cells mixed with Endothelial Cells. Sci. Rep. 2019, 9, 8920. [Google Scholar] [CrossRef] [Green Version]
- Pless, G.; Steffen, I.; Zeilinger, K.; Sauer, I.M.; Katenz, E.; Kehr, D.C.; Roth, S.; Mieder, T.; Schwartlander, R.; Müller, C.; et al. Evaluation of Primary Human Liver Cells in Bioreactor Cultures for Extracorporeal Liver Support on the Basis of Urea Production. Artif. Organs 2006, 30, 686–694. [Google Scholar] [CrossRef]
- Gerlach, J.C.; Mutig, K.; Sauer, I.M.; Schrade, P.; Efimova, E.; Mieder, T.; Naumann, G.; Grunwald, A.; Pless, G.; Mas, A.; et al. Use of primary human liver cells originating from discarded grafts in a bioreactor for liver support therapy and the prospects of culturing adult liver stem cells in bioreactors: A morphologic study. Transplantation 2003, 76, 781–786. [Google Scholar] [CrossRef]
- Lee-Montiel, F.T.; George, S.M.; Gough, A.H.; Sharma, A.D.; Wu, J.; DeBiasio, R.; Vernetti, L.A.; Taylor, D.L. Control of oxygen tension recapitulates zone-specific functions in human liver microphysiology systems. Exp. Biol. Med. 2017, 242, 1617–1632. [Google Scholar] [CrossRef] [Green Version]
- Gehre, C.; Flechner, M.; Kammerer, S.; Küpper, J.H.; Coleman, C.D.; Püschel, G.P.; Uhlig, K.; Duschl, C. Real time monitoring of oxygen uptake of hepatocytes in a microreactor using optical microsensors. Sci. Rep. 2020, 10, 13700. [Google Scholar] [CrossRef] [PubMed]
- Peel, S.; Corrigan, A.M.; Ehrhardt, B.; Jang, K.-J.; Caetano-Pinto, P.; Boeckeler, M.; Rubins, J.E.; Kodella, K.; Petropolis, D.B.; Ronxhi, J.; et al. Introducing an automated high content confocal imaging approach for Organs-on-Chips. Lab Chip 2019, 19, 410–421. [Google Scholar] [CrossRef] [PubMed]
- Bavli, D.; Prill, S.; Ezra, E.; Levy, G.; Cohen, M.; Vinken, M.; Vanfleteren, J.; Jaeger, M.; Nahmias, Y. Real-time monitoring of metabolic function in liver-on-chip microdevices tracks the dynamics of mitochondrial dysfunction. Proc. Natl. Acad. Sci. USA 2016, 113, E2231–E2240. [Google Scholar] [CrossRef] [Green Version]
- Prill, S.; Bavli, D.; Levy, G.; Ezra, E.; Schmälzlin, E.; Jaeger, M.S.; Schwarz, M.; Duschl, C.; Cohen, M.; Nahmias, Y. Real-time monitoring of oxygen uptake in hepatic bioreactor shows CYP450-independent mitochondrial toxicity of acetaminophen and amiodarone. Arch. Toxicol. 2016, 90, 1181–1191. [Google Scholar] [CrossRef] [PubMed]
- Hoffmann, S.A.; Müller-Vieira, U.; Biemel, K.; Knobeloch, D.; Heydel, S.; Lübberstedt, M.; Nüssler, A.K.; Andersson, T.B.; Gerlach, J.C.; Zeilinger, K. Analysis of drug metabolism activities in a miniaturized liver cell bioreactor for use in pharmacological studies. Biotechnol. Bioeng. 2012, 109, 3172–3181. [Google Scholar] [CrossRef] [PubMed]
- Zeilinger, K.; Schreiter, T.; Darnell, M.; Söderdahl, T.; Lübberstedt, M.; Dillner, B.; Knobeloch, D.; Nüssler, A.K.; Gerlach, J.C.; Andersson, T.B. Scaling down of a clinical three-dimensional perfusion multicompartment hollow fiber liver bioreactor developed for extracorporeal liver support to an analytical scale device useful for hepatic pharmacological in vitro studies. Tissue Eng. Part C Methods 2011, 17, 549–556. [Google Scholar] [CrossRef]
- Lübberstedt, M.; Müller-Vieira, U.; Biemel, K.M.; Darnell, M.; Hoffmann, S.A.; Knöspel, F.; Wönne, E.C.; Knobeloch, D.; Nüssler, A.K.; Gerlach, J.C.; et al. Serum-free culture of primary human hepatocytes in a miniaturized hollow-fibre membrane bioreactor for pharmacological in vitro studies. J. Tissue Eng. Regen. Med. 2015, 9, 1017–1026. [Google Scholar] [CrossRef] [PubMed]
- Jang, K.-J.; Otieno, M.A.; Ronxhi, J.; Lim, H.-K.; Ewart, L.; Kodella, K.R.; Petropolis, D.B.; Kulkarni, G.; Rubins, J.E.; Conegliano, D. Reproducing human and cross-species drug toxicities using a Liver-Chip. Sci. Transl. Med. 2019, 11. [Google Scholar] [CrossRef] [PubMed]
- Du, Y.; Li, N.; Yang, H.; Luo, C.; Gong, Y.; Tong, C.; Gao, Y.; Lü, S.; Long, M. Mimicking liver sinusoidal structures and functions using a 3D-configured microfluidic chip. Lab Chip 2017, 17, 782–794. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Choi, Y.Y.; Seok, J.I.; Kim, D.S. Flow-Based Three-Dimensional Co-Culture Model for Long-Term Hepatotoxicity Prediction. Micromachines 2019, 11, 36. [Google Scholar] [CrossRef] [Green Version]
- Knowlton, S.; Tasoglu, S. A bioprinted liver-on-a-chip for drug screening applications. Trends Biotechnol. 2016, 34, 681–682. [Google Scholar] [CrossRef]
- Ma, L.-D.; Wang, Y.-T.; Wang, J.-R.; Wu, J.-L.; Meng, X.-S.; Hu, P.; Mu, X.; Liang, Q.-L.; Luo, G.-A. Design and fabrication of a liver-on-a-chip platform for convenient, highly efficient, and safe in situ perfusion culture of 3D hepatic spheroids. Lab Chip 2018, 18, 2547–2562. [Google Scholar] [CrossRef]
- Corrado, B.; De Gregorio, V.; Imparato, G.; Attanasio, C.; Urciuolo, F.; Netti, P.A. A three-dimensional microfluidized liver system to assess hepatic drug metabolism and hepatotoxicity. Biotechnol. Bioeng. 2019, 116, 1152–1163. [Google Scholar] [CrossRef]
- Meng, Q.; Wang, Y.; Li, Y.; Shen, C. Hydrogel microfluidic-based liver-on-a-chip: Mimicking the mass transfer and structural features of liver. Biotechnol. Bioeng. 2021, 118, 612–621. [Google Scholar] [CrossRef]
- Hong, G.; Kim, J.; Oh, H.; Yun, S.; Kim, C.M.; Jeong, Y.M.; Yun, W.S.; Shim, J.H.; Jang, I.; Kim, C.Y.; et al. Production of Multiple Cell-Laden Microtissue Spheroids with a Biomimetic Hepatic-Lobule-Like Structure. Adv. Mater. 2021, e2102624. [Google Scholar] [CrossRef]
- Ulvestad, M.; Darnell, M.; Molden, E.; Ellis, E.; Åsberg, A.; Andersson, T.B. Evaluation of organic anion-transporting polypeptide 1B1 and CYP3A4 activities in primary human hepatocytes and HepaRG cells cultured in a dynamic three-dimensional bioreactor system. J. Pharmacol. Exp. Ther. 2012, 343, 145–156. [Google Scholar] [CrossRef] [Green Version]
- Boul, M.; Benzoubir, N.; Messina, A.; Ghasemi, R.; Mosbah, I.B.; Duclos-Vallée, J.C.; Dubart-Kupperschmitt, A.; Le Pioufle, B. A versatile microfluidic tool for the 3D culture of HepaRG cells seeded at various stages of differentiation. Sci. Rep. 2021, 11, 14075. [Google Scholar] [CrossRef] [PubMed]
- Jang, M.; Kleber, A.; Ruckelshausen, T.; Betzholz, R.; Manz, A. Differentiation of the human liver progenitor cell line (HepaRG) on a microfluidic-based biochip. J. Tissue Eng. Regen. Med. 2019, 13, 482–494. [Google Scholar] [CrossRef] [PubMed]
- Rennert, K.; Steinborn, S.; Gröger, M.; Ungerböck, B.; Jank, A.-M.; Ehgartner, J.; Nietzsche, S.; Dinger, J.; Kiehntopf, M.; Funke, H. A microfluidically perfused three dimensional human liver model. Biomaterials 2015, 71, 119–131. [Google Scholar] [CrossRef] [PubMed]
- Wang, Y.; Wang, H.; Deng, P.; Chen, W.; Guo, Y.; Tao, T.; Qin, J. In situ differentiation and generation of functional liver organoids from human iPSCs in a 3D perfusable chip system. Lab Chip 2018, 18, 3606–3616. [Google Scholar] [CrossRef] [PubMed]
- Bircsak, K.M.; DeBiasio, R.; Miedel, M.; Alsebahi, A.; Reddinger, R.; Saleh, A.; Shun, T.; Vernetti, L.A.; Gough, A. A 3D microfluidic liver model for high throughput compound toxicity screening in the OrganoPlate®. Toxicology 2021, 450, 152667. [Google Scholar] [CrossRef] [PubMed]
pHHs | HepG2 | HepaRG | iPSC-heps | Upcytes | |
---|---|---|---|---|---|
Availability | + | +++ | +++ | ++ | ++ |
Low costs | - | +++ | + | - | + |
Ease of use | ++ | +++ | + 1 | + 1 | ++ |
Proliferation/lifespan | - | +++ | +++ | +++ | ++ 2 |
Albumin synthesis | +++ | + | ++ | + | +++ |
Urea production | +++ | + | - | + | +++ |
Glycogen storage | +++ | +++ | +++ | +++ | +++ |
Phase I enzyme expression | +++ | - | ++ 3 | + 4 | ++ |
Phase II enzyme expression | +++ | - | +++ | + 4 | ++ |
Ease of Use | Long-Term Maintenance of Hepatic Properties | Suitability for DILI Prediction | Compatible with High Throughput Screening | |
---|---|---|---|---|
Spheroids | ||||
pHHs | ++ | +++ | +++ | ++ |
HepG2 | +++ | + | + | +++ |
HepaRG | + 1 | ++ | +++ | +++ |
iPSC-heps | + 1 | + 2 | + | ++ |
upcytes | ++ | n.d. | n.d. | ++ |
Perfused Bioreactors | ||||
pHHs | + | +++ | +++ | + |
HepG2 | ++ | + | n.d. | ++ |
HepaRG | + 1 | ++ | n.d. | ++ |
iPSC-heps | + 1 | + 2 | n.d. | + |
upcytes | + 3 | n.d. | n.d. | + 3 |
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Kammerer, S. Three-Dimensional Liver Culture Systems to Maintain Primary Hepatic Properties for Toxicological Analysis In Vitro. Int. J. Mol. Sci. 2021, 22, 10214. https://doi.org/10.3390/ijms221910214
Kammerer S. Three-Dimensional Liver Culture Systems to Maintain Primary Hepatic Properties for Toxicological Analysis In Vitro. International Journal of Molecular Sciences. 2021; 22(19):10214. https://doi.org/10.3390/ijms221910214
Chicago/Turabian StyleKammerer, Sarah. 2021. "Three-Dimensional Liver Culture Systems to Maintain Primary Hepatic Properties for Toxicological Analysis In Vitro" International Journal of Molecular Sciences 22, no. 19: 10214. https://doi.org/10.3390/ijms221910214