Evaluation of fibrosis in precision-cut tissue slices

Westra, Inge M.; Pham, Bao Tung; Groothuis, Geny M. M.; Olinga, Peter

doi:10.3109/00498254.2012.723151

Cited by 36 publications

(43 citation statements)

References 162 publications

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“…These effects were observed in PCLuS but not in PCKS. This could perhaps be caused by differences in the amount and type of immune cells present in both tissue types, as mentioned previously [6]. As these reductions are similar between non-targeting and Gapdh-targeting Accell siRNA, we hypothesize that Accell siRNA acts as an antagonist on siRNA sequence-independent proteins PKR and RIG-1, which are both present in the cytoplasm.…”

Section: Slice Inflammationmentioning

confidence: 86%

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siRNA-Mediated RNA Interference in Precision-Cut Tissue Slices Prepared from Mouse Lung and Kidney

Ruigrok

Maggan

Willaert

et al. 2017

AAPS J

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Abstract.Small interfering RNA (siRNA)-mediated RNAi interference (RNAi) is a powerful post-transcriptional gene silencing mechanism which can be used to study the function of genes in vitro (cell cultures) and in vivo (animal models). However, there is a translational gap between these models. Hence, there is a need for novel experimental models that combine the advantages of in vitro and in vivo models (e.g., simplicity, flexibility, throughput, and representability) to study the effects of siRNA. This need may be addressed by precision-cut tissue slices (PCTS), which represent an ex vivo model that mimics the structural and functional characteristics of a whole organ. The goal of this study was to investigate whether self-deliverable siRNA (Accell siRNA) can be used in precision-cut lung slices (PCLuS) and precision-cut kidney slices (PCKS) to achieve RNAi ex vivo. PCLuS and PCKS were prepared from mouse tissue, and they were subsequently incubated up to 48 h with no siRNA (untransfected), non-targeting Accell siRNA, or Gapdhtargeting Accell siRNA. Significant Gapdh mRNA silencing was achieved (PCLuS~55%; PCKS 40%) without compromising the viability and morphology of slices. Fluorescence microscopy confirmed that Accell siRNA diffused into PCLuS and PCKS. Spontaneous inflammation upon incubation was observed in PCLuS and PCKS as shown by a higher mRNA expression of proinflammatory cytokines Il1b, Il6, and Tnfa, although Accell siRNA appeared to diminish this response in PCLuS after 24 h. In conclusion, this ex vivo transfection model can be used to evaluate the effects of siRNA in relevant biological environments.

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Section: Slice Inflammationmentioning

confidence: 86%

“…PCTS refer to viable explants, with a welldefined thickness and diameter, prepared from tissue (e.g., lung and kidney) that can be cultured ex vivo [5,6]. In comparison to conventional in vitro models, PCTS offer crucial advantages.…”

Section: Introductionmentioning

confidence: 99%

siRNA-Mediated RNA Interference in Precision-Cut Tissue Slices Prepared from Mouse Lung and Kidney

Ruigrok

Maggan

Willaert

et al. 2017

AAPS J

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“…Different studies have been performed with brotic liver slices derived from animal models of chemicallyinduced brosis. 94 These studies focused on antibrotic therapy, 105,106 the detection of biomarkers and the mechanism of brosis. 107,108 Veidal et al 107 showed that a specic fragment of type III collagen generated by metalloproteinase-2 and metalloproteinase-9 by brotic liver slices is a biomarker for liver brosis.…”

Section: Precision-cut Liver Slices In Liver Fibrosismentioning

confidence: 99%

“…PCTS represent an ex vivo tissue culture technique that replicates most of the multicellular characteristics of whole organs in vivo. 94 Recently, the utilisation of PCTS for the study of organ brosis has been reviewed. 94 Particularly, the possibility of using (diseased) human tissue to study mechanisms of brosis has the potential to become a great asset of the PCTS technique.…”

Section: Fibrosis In Liver and Intestinementioning

confidence: 99%

“…94 Recently, the utilisation of PCTS for the study of organ brosis has been reviewed. 94 Particularly, the possibility of using (diseased) human tissue to study mechanisms of brosis has the potential to become a great asset of the PCTS technique. Organs share major common disease pathways, but for immune cells and especially myobroblasts, the main extracellular matrix producing cells, there are organ-and aetiology-specic variations in cellular gene expression programmes.…”

Section: Fibrosis In Liver and Intestinementioning

confidence: 99%

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Translational Research in Pharmacology and Toxicology Using Precision-Cut Tissue Slices

Groothuis

Casini

Olinga

2014

Human-Based Systems for Translational Research

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In this chapter we discuss the application of human liver, intestine, lung and tumour precision-cut tissue slices (PCTS) as a translational ex vivo model in studies on ADME (absorption, distribution, metabolism and excretion) and toxicology of drugs, and for studies on diseases such as fibrosis in the liver and the intestine, obstructive lung diseases, viral infections and cancer. As the use of PCTS in research is steadily increasing it is impossible to give a fully comprehensive review of all applications of PCTS, but by highlighting some of the most important examples with a special emphasis on the application of human PCTS, we aim to show the extensive potential of this versatile technique in pathology and drug research.

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A Bioreactor Technology for Modeling Fibrosis in Human and Rodent Precision‐Cut Liver Slices

et al. 2019

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Precision cut liver slices (PCLSs) retain the structure and cellular composition of the native liver and represent an improved system to study liver fibrosis compared to two‐dimensional mono‐ or co‐cultures. The aim of this study was to develop a bioreactor system to increase the healthy life span of PCLSs and model fibrogenesis. PCLSs were generated from normal rat or human liver, or fibrotic rat liver, and cultured in our bioreactor. PCLS function was quantified by albumin enzyme‐linked immunosorbent assay (ELISA). Fibrosis was induced in PCLSs by transforming growth factor beta 1 (TGFβ1) and platelet‐derived growth factor (PDGFββ) stimulation ± therapy. Fibrosis was assessed by gene expression, picrosirius red, and α‐smooth muscle actin staining, hydroxyproline assay, and soluble ELISAs. Bioreactor‐cultured PCLSs are viable, maintaining tissue structure, metabolic activity, and stable albumin secretion for up to 6 days under normoxic culture conditions. Conversely, standard static transwell‐cultured PCLSs rapidly deteriorate, and albumin secretion is significantly impaired by 48 hours. TGFβ1/PDGFββ stimulation of rat or human PCLSs induced fibrogenic gene expression, release of extracellular matrix proteins, activation of hepatic myofibroblasts, and histological fibrosis. Fibrogenesis slowly progresses over 6 days in cultured fibrotic rat PCLSs without exogenous challenge. Activin receptor‐like kinase 5 (Alk5) inhibitor (Alk5i), nintedanib, and obeticholic acid therapy limited fibrogenesis in TGFβ1/PDGFββ‐stimulated PCLSs, and Alk5i blunted progression of fibrosis in fibrotic PCLS. Conclusion: We describe a bioreactor technology that maintains functional PCLS cultures for 6 days. Bioreactor‐cultured PCLSs can be successfully used to model fibrogenesis and demonstrate efficacy of antifibrotic therapies.

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Evaluation of fibrosis in precision-cut tissue slices

Cited by 36 publications

References 162 publications

siRNA-Mediated RNA Interference in Precision-Cut Tissue Slices Prepared from Mouse Lung and Kidney

siRNA-Mediated RNA Interference in Precision-Cut Tissue Slices Prepared from Mouse Lung and Kidney

Translational Research in Pharmacology and Toxicology Using Precision-Cut Tissue Slices

A Bioreactor Technology for Modeling Fibrosis in Human and Rodent Precision‐Cut Liver Slices

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