Preparation and incubation of precision-cut liver and intestinal slices for application in drug metabolism and toxicity studies

Graaf, Inge A. M. de; Olinga, Peter; Jager, Marina H. de; Merema, Marjolijn T.; Kanter, Ruben de; Kerkhof, Esther G. van de; Groothuis, Geny M. M.

doi:10.1038/nprot.2010.111

Cited by 342 publications

(461 citation statements)

References 97 publications

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“…In contrast to isolated hepatocytes and hepatocarcinoma cell lines, Precision Cut Liver Slices (PCLS) represent a multicellular ex vivo model for the investigation of drug induced liver toxicology. Using optimized culture conditions, the viability and organ-specific architectural composition of the different cell types within rat PCLS can be maintained up to 96 h in culture [5,6]. However, comparable with observations in hepatocytes, the expression of genes from multiple metabolic pathways such as carbohydrate metabolism and lipid metabolism is altered in PCLS that are kept in culture for 24 h or longer [6,7].…”

Section: Introductionmentioning

confidence: 87%

“…Thus, the rapidly altered gene expression of PCLS upon cultivation restricts their use to short-term applications and does not allow the investigation of drug-induced subchronic toxicity [5,11]. In addition to their restricted metabolic activity during prolonged cultivation, a major limitation of PCLS is the requirement of complex and sophisticated dynamic culture conditions, preventing high-throughput screening applications [5].…”

Section: Introductionmentioning

confidence: 99%

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Characterization of a Rat Multi-Cell Type 3D-Liver Microtissue System

Odermatt¹

2013

J Tissue Sci Eng

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Background: Primary hepatocytes rapidly lose their polarized morphology and the expression of important liverspecific metabolic enzymes, receptors and transport proteins under normal two-dimensional (2D) culture conditions. Thus, their use as a reliable predictive in vitro model for drug-induced liver injury is limited. Three-dimensional (3D) liver micro tissue culture systems have become an increasingly attractive alternative for the evaluation of druginduced liver toxicity. However, several liver-specific pathways remain to be characterized in such models. Methods and principal findings:In the present work, we compared the expression of several genes with a role in the anti-oxidant cell defense and glucocorticoid pathways in rat H4IIE hepatoma cells, primary rat hepatocytes, 2D-hepatocyte sandwich culture, and a multi-cell type micro tissue model comprising primary rat hepatocytes in coculture with liver-derived nonparenchymal cells and macrophage (Kupffer cells). Gene expression was studied for up to 25 days in culture. High expression levels of the Nrf2-dependent genes NQO1, ABCC3 and GST2A were detected in H4IIE cells and in 3D-liver microtissues, in contrast to 2D-hepatocytes where a rapid decline of these genes was observed. The glucocorticoid-dependent genes ORM1, G6PC, PCK1 and HSD11B1 were highly expressed up to 25 days of cultivation in 3D-liver microtissues, but they showed very low or background expression in H4IIE and 2D-hepatocyte models. Conclusions:The tested 3D-multi-cell type liver micro tissue represents a stable and functionally active model system, with sustained expression for more than three weeks of cultivation of important metabolic proteins regulated by the glucocorticoid and anti-oxidant cell defense pathway.

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Section: Introductionmentioning

confidence: 87%

Section: Introductionmentioning

confidence: 99%

Characterization of a Rat Multi-Cell Type 3D-Liver Microtissue System

Odermatt¹

2013

J Tissue Sci Eng

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“…They closely resemble the organ from which it is prepared, with all cell types present in their original tissue-matrix configuration (78). The circumstance that in tissue slices the architecture of the original organ is retained makes them an attractive tool for drug delivery studies.…”

Section: Issues To Be Addressed For Further Development Of Ec Specifimentioning

confidence: 99%

Targeted siRNA delivery to diseased microvascular endothelial cells—Cellular and molecular concepts

et al. 2011

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SummaryIncreased insight in the role of endothelial cells in the pathophysiology of cancer, inflammatory and cardiovascular diseases, has drawn great interest in pharmacological interventions aiming at the endothelium in diseased sites. Their location in the body makes them suitable targets for therapeutic approaches based on targeted drug delivery. Functional heterogeneity of the microvascular bed in normal organ homeostasis has been appreciated for a long time, and more recent studies have revealed heterogeneity in endothelial reactivity to inflammatory stimuli as well. Upon stimulation, each organ displays a vascular bed specific pattern of cell adhesion molecules providing challenging opportunities to deliver drugs or small RNAs to organ specific (micro)vascular endothelial subsets. In this review we introduce general concepts of endothelial heterogeneity in relation to disease state and its consequences for targeted therapeutic interventions. Furthermore, we will describe novel approaches to interfere with endothelial cell engagement in disease with a main focus on siRNA therapeutics and currently used nonviral lipid and polymer-based siRNA delivery systems. The last part of this review addresses some technical issues that are essential in proving the concept of target mRNA knock down in a vascular bed specific manner, and the further development of effective endothelial cell specific drug delivery devices.

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“…A promising alternative approach that provides a better in vivo-like environment is the use of precision-cut tissue slices (PCS), which represent an ex vivo model of the organ of study by maintaining the original architecture, i.e., containing all the cells types of the tissue in their natural conformation (42). The advantage of this system is that slices from different species can be prepared and compared, such as from rodents as well as human biopsy material.…”

Section: Ex Vivo Tissue Preparation: Bringing In Vivo Tissue Into Petmentioning

confidence: 99%

In vitro-ex vivo model systems for nanosafety assessment

Wick

Chortarea

Guénat³

et al. 2015

European Journal of Nanomedicine

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Engineered nanomaterials have unique and novel properties enabling wide-ranging new applications in nearly all fields of research. As these new properties have raised concerns about potential adverse effects for the environment and human health, extensive efforts are underway to define reliable, cost-and time-effective, as well as mechanistic-based testing strategies to replace the current method of animal testing, which is still the most prevalent model used for the risk assessment of chemicals. Current approaches for nanomaterials follow this line. The aim of this review is to explore and qualify the relevance of new in vitro and ex vivo models in (nano) material safety assessment, a crucial prerequisite for translation into applications.

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Preparation and incubation of precision-cut liver and intestinal slices for application in drug metabolism and toxicity studies

Cited by 342 publications

References 97 publications

Characterization of a Rat Multi-Cell Type 3D-Liver Microtissue System

Characterization of a Rat Multi-Cell Type 3D-Liver Microtissue System

Targeted siRNA delivery to diseased microvascular endothelial cells—Cellular and molecular concepts

In vitro-ex vivo model systems for nanosafety assessment

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