Comparison of Hepatic 2D Sandwich Cultures and 3D Spheroids for Long-term Toxicity Applications: A Multicenter Study

Toxicological Sciences, Journal Year: 2018, Volume and Issue: 162(2), P. 655 - 666

Published: Jan. 9, 2018

Primary human hepatocytes (PHHs) are commonly used for in vitro studies of drug-induced liver injury. However, when cultured as 2D monolayers, PHH lose crucial hepatic functions within hours. This dedifferentiation can be ameliorated PHHs sandwich configuration (2Dsw), particularly cultures regularly re-overlaid with extracellular matrix, or 3D spheroids. In this study, the 6 participating laboratories evaluated robustness these 2 model systems made from cryopreserved same donors considering both inter-donor and inter-laboratory variability compared their suitability use repeated-dose toxicity using 5 different hepatotoxins mechanisms. We found that expression levels proteins involved drug absorption, distribution, metabolism, excretion, well catalytic activities CYPs, were significantly higher spheroid cultures, potentially affecting exposure cells to drugs metabolites. Furthermore, global proteomic analyses revealed temporally stable whereas proteomes 2Dsw showed substantial alterations protein patterns over 14 days culture. Overall, more sensitive hepatotoxic compounds investigated, upon long-term exposures, across testing sites little variability. The data presented here suggest repeated-dosing regimens improve predictivity assays, spheroids provide a robust system mechanistic hepatotoxicity, has dedifferentiated phenotype lower sensitivity detect hepatotoxicity.

Language: Английский

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Engineered Liver-On-A-Chip Platform to Mimic Liver Functions and Its Biomedical Applications: A Review

Micromachines, Journal Year: 2019, Volume and Issue: 10(10), P. 676 - 676

Published: Oct. 7, 2019

Hepatology and drug development for liver diseases require in vitro models. Typical models include 2D planar primary hepatocytes, hepatocyte spheroids, organoids, liver-on-a-chip. Liver-on-a-chip has emerged as the mainstream model because it recapitulates microenvironment good assay robustness such reproducibility. with human cells can potentially correlate clinical testing. not only predict hepatotoxicity metabolism, but also connect other artificial organs on chip a human-on-a-chip, which reflect overall effect of drug. Engineering an effective liver-on-a-chip device requires knowledge multiple disciplines including chemistry, fluidic mechanics, cell biology, electrics, optics. This review first introduces physiological microenvironments liver, especially composition its specialized roles, then summarizes strategies to build via microfluidic technologies biomedical applications. In addition, latest advancements are discussed, serve basis further research.

Language: Английский

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Managing the challenge of drug-induced liver injury: a roadmap for the development and deployment of preclinical predictive models

Nature Reviews Drug Discovery, Journal Year: 2019, Volume and Issue: 19(2), P. 131 - 148

Published: Nov. 20, 2019

Language: Английский

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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 Metabolism and Disposition, Journal Year: 2017, Volume and Issue: 45(4), P. 419 - 429

Published: Jan. 30, 2017

Reliable and versatile hepatic in vitro systems for the prediction of drug pharmacokinetics toxicity are essential constituents preclinical safety assessment pipelines new medicines. Here, we compared three emerging cell systems-hepatocytes derived from induced pluripotent stem cells, HepaRG three-dimensional primary human hepatocyte (PHH) spheroids-at transcriptional functional levels a multicenter study to evaluate their potential as predictive models drug-induced hepatotoxicity. Transcriptomic analyses revealed widespread gene expression differences between models, with 8148 17,462 analyzed genes (47%) being differentially expressed. Expression involved metabolism endogenous well xenobiotic compounds were significantly elevated PHH spheroids, whereas division endocytosis upregulated cells hepatocytes respectively. Consequently, spheroids more sensitive panel drugs distinctly different mechanisms, an effect that was amplified by long-term exposure using repeated treatments. Importantly, toxicogenomic transcriptomic changes compliance cholestatic, carcinogenic, or steatogenic vivo mechanisms at clinically relevant concentrations. Combined, data reveal important phenotypic suggest can be used investigations liver injury humans.

Language: Английский

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Prediction of Drug-Induced Hepatotoxicity Using Long-Term Stable Primary Hepatic 3D Spheroid Cultures in Chemically Defined Conditions

Toxicological Sciences, Journal Year: 2018, Volume and Issue: 163(2), P. 655 - 665

Published: March 23, 2018

High failure rates of drug candidates in the clinics, restricted-use warnings as well withdrawals drugs postmarketing stages are substantial concern for pharmaceutical industry and drug-induced liver injury (DILI) constitutes one most frequent reasons such safety failures. Importantly, DILI cannot be accurately predicted using animal models, tests commonly complemented with assessments human vitro systems. 3D spheroid cultures primary hepatocytes chemically defined conditions, hereafter termed CD-spheroids, have recently emerged a microphysiological model system which retain their molecular phenotypes hepatic functions multiple weeks culture. However, predictive power detection hepatotoxic liabilities has not been systematically assessed. Therefore, we here evaluated hepatotoxicity 123 or without direct implication clinical events. ATP quantifications single endpoint, distinguished between nontoxic structural analogues exceeded both sensitivity specificity all previously published assays at substantially lower exposure levels, successfully detecting 69% compounds producing any false positive results (100% specificity). Furthermore, platform supports culture spheroids from preclinical thereby allowing identification animal-specific toxicity We anticipate that CD-spheroids represent powerful versatile tool discovery development can reliably flag provide guidance selection suitable models.

Language: Английский

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Endogenous and xenobiotic metabolic stability of primary human hepatocytes in long‐term 3D spheroid cultures revealed by a combination of targeted and untargeted metabolomics

The FASEB Journal, Journal Year: 2017, Volume and Issue: 31(6), P. 2696 - 2708

Published: March 6, 2017

Adverse reactions or lack of response to medications are important concerns for drug development programs. However, faithful predictions metabolism and toxicity difficult because animal models show only limited translatability humans. Furthermore, current in vitro systems, such as hepatic cell lines primary human hepatocyte (PHH) 2-dimensional (2D) monolayer cultures, can be used acute tests their immature phenotypes inherent instability. Therefore, the migration novel phenotypically stable is prime importance pharmaceutical industry. Novel 3-dimensional (3D) culture systems have been shown accurately mimic vivo on transcriptomic proteomic level, but information about metabolic stability lacking. Using a combination targeted untargeted high-resolution mass spectrometry, we found that PHHs 3D spheroid cultures remained metabolically multiple weeks, whereas patterns from same donors cultured conventional 2D monolayers rapidly deteriorated. pharmacokinetic differences between were maintained enabling studies interindividual variability toxicity. We conclude system constitutes suitable model long-term pharmacokinetics.-Vorrink, S. U., Ullah, S., Schmid, Nandania, J., Velagapudi, V., Beck, O., Ingelman-Sundberg, M., Lauschke, V. M. Endogenous xenobiotic hepatocytes revealed by metabolomics.

Language: Английский

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Engineered cell and tissue models of pulmonary fibrosis

Advanced Drug Delivery Reviews, Journal Year: 2017, Volume and Issue: 129, P. 78 - 94

Published: Dec. 18, 2017

Language: Английский

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Three‐dimensional liver‐derived extracellular matrix hydrogel promotes liver organoids function

Journal of Cellular Biochemistry, Journal Year: 2017, Volume and Issue: 119(6), P. 4320 - 4333

Published: Dec. 16, 2017

An important advantage of employing extracellular matrix (ECM)-derived biomaterials in tissue engineering is the ability to tailor biochemical and biophysical microenvironment cells. This study aims assess whether three-dimensional (3D) liver-derived ECM hydrogel (LEMgel) promotes physiological function liver organoids generated by self-organization human hepatocarcinoma cells together with mesenchymal endothelial We have optimized decellularization method fabricate derived from sheep preserve greatest content glycosaminoglycans, collagen, laminin, fibronectin produced LEMgel. During gelation, complex viscoelasticity modulus LEMgel (3 mg/mL) increased 186.7 1570.5 Pa Tan Delta decreased 0.27 0.18. Scanning electron microscopy (SEM) determined that had a pore size 382 ± 71 µm. Hepatocarcinoma self-organized 3D (LEMgel organoids) showed an epithelial phenotype expressed ALB, CYP3A4, E-cadherin, ASGPR. The organoid significant upregulation transcripts CYP3A7, TAT as well downregulation AFP compared collagen type I- hydrogel-free-organoids or solubilized LEM 2D culture Generated significantly more ALB AAT secretion, urea production, CYP3A4 enzyme activity, inducibility. In conclusion, enhanced functional activity traditional 2D, 3D, gel cultures. Our novel could potentially be used engineering, drug discovery, toxicology studies, bio-artificial fabrication.

Language: Английский

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Generation of a 3D Liver Model Comprising Human Extracellular Matrix in an Alginate/Gelatin-Based Bioink by Extrusion Bioprinting for Infection and Transduction Studies

International Journal of Molecular Sciences, Journal Year: 2018, Volume and Issue: 19(10), P. 3129 - 3129

Published: Oct. 12, 2018

Bioprinting is a novel technology that may help to overcome limitations associated with two-dimensional (2D) cell cultures and animal experiments, as it allows the production of three-dimensional (3D) tissue models composed human cells. The present study describes optimization bioink alginate, gelatin extracellular matrix (hECM) print HepaRG liver cells pneumatic extrusion printer. resulting model was tested for its suitability transduction by an adeno-associated virus (AAV) vector infection adenovirus 5 (hAdV5). We found supplementation basic alginate/gelatin 0.5 1 mg/mL hECM provides desirable properties printing process, stability printed constructs, viability metabolic functions were efficiently transduced AAV vectors serotype 6, which successfully silenced endogenous target (cyclophilin B) means RNA interference. Furthermore, 3D supported efficient adenoviral replication making suitable biology develop new antiviral compounds. consider approach described here paradigmatic development studies including viral infectious viruses.

Language: Английский

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3D Primary Hepatocyte Culture Systems for Analyses of Liver Diseases, Drug Metabolism, and Toxicity: Emerging Culture Paradigms and Applications

Biotechnology Journal, Journal Year: 2019, Volume and Issue: 14(7)

Published: April 8, 2019

Recent research has shown that the maintenance of relevant liver functions ex vivo requires models in which cells exhibit an vivo-like phenotype, often achieved by reconstitution appropriate cellular interactions. Multiple different have been presented differ utilized, media, and culture conditions. Furthermore, several technologically approaches including bioreactors, chips, plate-based systems fluidic or static media constituting chemically diverse materials. Using such models, ability to predict drug metabolism, toxicity, functionality increased tremendously as compared conventional vitro are cultured 2D monolayers. Here, authors highlight important considerations for microphysiological primary hepatocyte culture, review current paradigms, discuss their opportunities studies hepatotoxicity, biology, disease.

Language: Английский

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