Engineering in vitro models of hepatofibrogenesis

Mazza, Giuseppe; Al‐Akkad, Walid; Rombouts, Krista

doi:10.1016/j.addr.2017.05.018

Cited by 49 publications

(47 citation statements)

References 192 publications

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“…Over the past decade, several studies have demonstrated the appropriateness of using naturally occurring ECM scaffolds derived by decellularized human or animal tissues for tissue engineering. In this context, liver bioengineering could be used for transplantation44 and for drug toxicity testing in 3D in vitro cultures 45. There is convincing experimental evidence that decellularization–recellularization technologies provide a valuable platform for liver bioengineering through the repopulation of liver ECM scaffolds with parenchymal and nonparenchymal liver cells, thus recapitulating, at least in part, natural tissue complexity.…”

Section: Decellularized 3d Ecm Scaffolds For Tissue/whole Organ Enginmentioning

confidence: 99%

Liver tissue engineering: From implantable tissue to whole organ engineering

Mazza

Al‐Akkad

Rombouts

et al. 2017

Hepatology Communications

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101

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The term “liver tissue engineering” summarizes one of the ultimate goals of modern biotechnology: the possibility of reproducing in total or in part the functions of the liver in order to treat acute or chronic liver disorders and, ultimately, create a fully functional organ to be transplanted or used as an extracorporeal device. All the technical approaches in the area of liver tissue engineering are based on allocating adult hepatocytes or stem cell‐derived hepatocyte‐like cells within a three‐dimensional structure able to ensure their survival and to maintain their functional phenotype. The hosting structure can be a construct in which hepatocytes are embedded in alginate and/or gelatin or are seeded in a pre‐arranged scaffold made with different types of biomaterials. According to a more advanced methodology termed three‐dimensional bioprinting, hepatocytes are mixed with a bio‐ink and the mixture is printed in different forms, such as tissue‐like layers or spheroids. In the last decade, efforts to engineer a cell microenvironment recapitulating the dynamic native extracellular matrix have become increasingly successful, leading to the hope of satisfying the clinical demand for tissue (or organ) repair and replacement within a reasonable timeframe. Indeed, the preclinical work performed in recent years has shown promising results, and the advancement in the biotechnology of bioreactors, ex vivo perfusion machines, and cell expansion systems associated with a better understanding of liver development and the extracellular matrix microenvironment will facilitate and expedite the translation to technical applications. (Hepatology Communications 2018;2:131–141)

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Section: Decellularized 3d Ecm Scaffolds For Tissue/whole Organ Enginmentioning

confidence: 99%

Liver tissue engineering: From implantable tissue to whole organ engineering

Mazza

Al‐Akkad

Rombouts

et al. 2017

Hepatology Communications

Self Cite

101

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“…Currently, liver cell mono or co-cultures, sandwich cultures, organoids or animal models are used to interrogate the mechanisms driving pathogenesis and reversion of liver disease and fibrosis to identify new targetable pathways and test anti-fibrotic drugs[2-4]. These preclinical tools have strengths but also limitations.…”

Section: Introductionmentioning

confidence: 99%

“…Hepatocytes rapidly dedifferentiate and down-regulate synthesis of albumin, metabolic enzymes and cytochrome P450 after 24 hours in culture[7]. 3D spheroids provide an alternative system to grow hepatocytes or mixed liver cell cultures but they fail to recapitulate the structural organisation of the liver or maintain physiologically relevant interactions with the ECM[4].…”

Section: Introductionmentioning

confidence: 99%

A novel bioreactor technology for modelling fibrosis in human and rodent precision-cut liver slices

Paish

Reed

Brown

et al. 2018

Preprint

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Summary boxWhat is already known about this subject?Currently there are no effective anti-fibrotic drugs to treat liver fibrosis and there is an urgent unmet need to increase our knowledge of the disease process and develop better tools for anti-fibrotic drug discovery.Preclinical in vitro cell cultures and animal models are widely used to study liver fibrosis and test anti-fibrotic drugs, but have shortfalls; cell culture models lack the relevant complex cell-cell interactions of the liver and animal models only reproduce some features of human disease.Precision Cut Liver Slices (PCLS) are structurally representative of the liver and can be used to model liver fibrosis and test anti-fibrotic drugs. However, PCLS are typically cultured in elevated, non-physiological oxygen levels and only have a healthy lifespan of 48h.What are the new findings?We have developed a novel bioreactor culture system that increases the longevity of functional PCLS to up to 6 days under normoxic conditions.Bioreactor cultured PCLS can be used to model fibrogenesis in both normal and fibrotic PCLS using a combination of biochemical and histological outputs.Administration of an Alk5 inhibitor effectively limits fibrogenesis in normal rodent and human PCLS and in rodent PCLS with established fibrosis.How might it impact on clinical practice in the foreseeable future?The extended longevity of bioreactor cultured PCLS represent a novel pre-clinical tool to investigate the cellular and molecular mechanisms of liver fibrosis.Bioreactor cultured human PCLS offer a clinically relevant system to test efficacy of anti-fibrotic drugs.AbstractObjectivePrecision cut liver slices (PCLS) 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 objective of this study was to develop a bioreactor system to increase the healthy lifespan of PCLS and model fibrogenesis.DesignPCLS were generated from normal rat or human liver, or 4-week carbon tetrachloride-fibrotic rat liver and cultured in our patented bioreactor. PCLS function was quantified by albumin ELISA. Fibrosis was induced in PCLS by TGFβ1 and PDGFββ stimulation. Alk5 inhibitor therapy was used. Fibrosis was assessed by fibrogenic gene expression, Picrosirius Red and αSmooth Muscle Actin staining, hydroxyproline assay and collagen 1a1, fibronectin and hyaluronic acid ELISA.ResultsBioreactor cultured PCLS are viable, maintaining tissue structure and stable albumin secretion for up to 6 days under normoxic culture conditions. Conversely, standard static transwell cultured PCLS rapidly deteriorate and albumin secretion is significantly impaired by 48 hours. TGFβ1 and PDGFββ stimulation of rat or human PCLS 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 PCLS without exogenous challenge. Alk5 inhibitor limited fibrogenesis in both TGFβ1 and PDGFββ stimulated PCLS and fibrotic PCLS.ConclusionWe describe a new bioreactor technology which maintains functional PCLS cultures for 6 days. Bioreactor cultured PCLS can be successfully used to model fibrogenesis and demonstrate efficacy of an anti-fibrotic therapy.

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“…Currently, mono‐ or co‐cultures of liver cells, sandwich cultures, organoids, or animal models are used to interrogate the mechanisms driving the pathogenesis or reversion of liver disease and fibrosis to identify new targetable pathways and test antifibrotic drugs . These preclinical tools have strengths, but also limitations.…”

mentioning

confidence: 99%

“…Hepatocytes rapidly dedifferentiate and down‐regulate synthesis of albumin, metabolic enzymes, and cytochrome P450 after 24 hours in culture . Three‐dimensional (3D) spheroids provide an alternative system to grow hepatocytes or mixed liver cell cultures, but they fail to recapitulate the structural organisation of the liver or maintain physiologically relevant interactions with the ECM …”

mentioning

confidence: 99%

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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Engineering in vitro models of hepatofibrogenesis

Cited by 49 publications

References 192 publications

Liver tissue engineering: From implantable tissue to whole organ engineering

Liver tissue engineering: From implantable tissue to whole organ engineering

A novel bioreactor technology for modelling fibrosis in human and rodent precision-cut liver slices

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

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