Best Practices and Progress in Precision-Cut Liver Slice Cultures

Dewyse, Liza; Reynaert, Hendrik; Grunsven, Leo A. van

doi:10.3390/ijms22137137

Cited by 35 publications

(44 citation statements)

References 116 publications

(158 reference statements)

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“…To study MAFLD, the PCLS model is a promising tool that recapitulates the multicellular architecture of the liver. This technique supports the 3Rs of Replacement, Reduction and Refinement of animal use [46,47], since multiple conditions can be tested in the same liver. In this paper, steatosis was first induced in healthy murine liver slices.…”

Section: Discussionsupporting

confidence: 55%

The Effects of Butyrate on Induced Metabolic-Associated Fatty Liver Disease in Precision-Cut Liver Slices

et al. 2021

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Metabolic-associated fatty liver disease (MAFLD) starts with hepatic triglyceride accumulation (steatosis) and can progress to more severe stages such as non-alcoholic steatohepatitis (NASH) and even cirrhosis. Butyrate, and butyrate-producing bacteria, have been suggested to reduce liver steatosis directly and systemically by increasing liver β-oxidation. This study aimed to examine the influence of butyrate directly on the liver in an ex vivo induced MAFLD model. To maintain essential intercellular interactions, precision-cut liver slices (PCLSs) were used. These PCLSs were prepared from male C57BL/6J mice and cultured in varying concentrations of fructose, insulin, palmitic acid and oleic acid, to mimic metabolic syndrome. Dose-dependent triglyceride accumulation was measured after 24 and 48 h of incubation with the different medium compositions. PCLSs viability, as indicated by ATP content, was not affected by medium composition or the butyrate concentration used. Under induced steatotic conditions, butyrate did not prevent triglyceride accumulation. Moreover, it lowered the expression of genes encoding for fatty acid oxidation and only increased C4 related carnitines, which indicate butyrate oxidation. Nevertheless, butyrate lowered the fibrotic response of PCLSs, as shown by reduced gene expression of fibronectin, alpha-smooth muscle actin and osteopontin, and protein levels of type I collagen. These results suggest that in the liver, butyrate alone does not increase lipid β-oxidation directly but might aid in the prevention of MAFLD progression to NASH and cirrhosis.

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Section: Discussionsupporting

confidence: 55%

The Effects of Butyrate on Induced Metabolic-Associated Fatty Liver Disease in Precision-Cut Liver Slices

et al. 2021

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“…Several studies have evaluated the anti-fibrotic effects of drugs and direct activation of HSCs by TGFβ or/and PDGF-BB can be easily induced ( 16 ). However, the evaluation of drug-induced liver fibrosis and HSC activation has not been well documented yet ( 10 ). We set out to evaluate whether it is possible to induce HSC activation in traditional PCLS cultures by exposing them to acetaminophen (APAP), shown to transiently induce HSC activation after an acute exposure ( 24 ), and fibrosis when chronically administered to mice ( 25 ).…”

Section: Resultsmentioning

confidence: 99%

“…PCLS cultures are a very useful tool in metabolism studies and the testing of anti-fibrotic drugs ( 12 , 17 , 18 ), however drug-induced liver injury dependent HSC activation and fibrosis is poorly documented in PCLS cultures ( 10 ). In this study, we aimed at establishing culture conditions that would allow the induction of HSC activation depending on drug-induced liver damage instead of only depending on the damage of the slices that is intrinsic to the PCLS procedure.…”

Section: Discussionmentioning

confidence: 99%

“…Precision-cut liver slices (PCLS) were introduced 35 years ago as an in vitro liver model with great potential as it possesses all needed requirements. To obtain these cultures, fresh liver tissue is processed to yield 250 μm slices with a diameter of 3–8 mm that can be cultured in regular tissue culture plates ( 10 ). As the PCLS preparation results in cut surfaces which triggers a repair and regenerative response and thus activate the HSCs within 48 h ( 11 ), PCLS have been used as a model to test anti-fibrotic drugs ( 12 – 14 ).…”

Section: Introductionmentioning

confidence: 99%

“…Even though several researchers have tried to prolong the PCLS culture lifespan ( 15 , 16 ), PCLS are mostly used during a short period of time (24–48 h). While many (anti-)fibrotics have been tested ( 12 , 17 , 18 ), demonstrations of drug-induced hepatocyte-damage dependent HSC activation and fibrosis are lacking for PCLS culture set-ups ( 10 ).…”

Section: Introductionmentioning

confidence: 99%

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Improved Precision-Cut Liver Slice Cultures for Testing Drug-Induced Liver Fibrosis

et al. 2022

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In vitro models of human liver disease often fail to mimic the complex 3D structures and cellular organizations found in vivo. Precision cut liver slices (PCLS) retain the complex physiological architecture of the native liver and therefore could be an exceptional in vitro liver model. However, the production of PCLS induces a spontaneous culture-induced fibrogenic reaction, limiting the application of PCLS to anti-fibrotic compounds. Our aim was to improve PCLS cultures to allow compound-induced fibrosis induction. Hepatotoxicity in PCLS cultures was analyzed by lactate dehydrogenase leakage and albumin secretion, while fibrogenesis was analyzed by qRT-PCR and western blot for hepatic stellate cell (HSC) activation markers and collagen 6 secretion by enzyme-linked immunosorbent assays (ELISA). We demonstrate that supplementation of 3 mm mouse PCLS cultures with valproate strongly reduces fibrosis and improves cell viability in our PCLS cultures for up to 5 days. Fibrogenesis can still be induced both directly and indirectly through exposure to TGFβ and the hepatotoxin acetaminophen, respectively. Finally, human PCLS cultures showed similar but less robust results. In conclusion, we optimized PCLS cultures to allow for drug-induced liver fibrosis modeling.

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A Microfluidic Device for Long‐Term Maintenance of Organotypic Liver Cultures

Hoyos-Vega

Hong

Loutherback

et al. 2022

Adv Materials Technologies

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Liver cultures may be used for disease modeling, testing therapies, and predicting drug‐induced injury. The complexity of the liver cultures has evolved from hepatocyte monocultures to co‐cultures with non‐parenchymal cells and finally to precision‐cut liver slices. The latter culture format retains liver's native biomolecular and cellular complexity and therefore holds considerable promise for in vitro testing. However, liver slices remain functional for ≈72 h in vitro and display limited utility for some disease modeling and therapy testing applications that require longer culture times. This paper describes a microfluidic device for longer‐term maintenance of functional organotypic liver cultures. This microfluidic culture system is designed to enable direct injection of liver tissue into a culture chamber through a valve‐enabled side port. Liver tissue is embedded in collagen and remained functional for up to 31 days, highlighted by continued production of albumin and urea. These organotypic cultures also express several enzymes involved in xenobiotic metabolism. Conversely, matched liver tissue embedded in collagen in a 96‐well plate loses its phenotype and function within 3–5 days. The microfluidic organotypic liver cultures described here represent a significant advance in liver cultivation and may be used for future modeling of liver diseases or for individualized liver‐directed therapies.

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Best Practices and Progress in Precision-Cut Liver Slice Cultures

Cited by 35 publications

References 116 publications

The Effects of Butyrate on Induced Metabolic-Associated Fatty Liver Disease in Precision-Cut Liver Slices

The Effects of Butyrate on Induced Metabolic-Associated Fatty Liver Disease in Precision-Cut Liver Slices

Improved Precision-Cut Liver Slice Cultures for Testing Drug-Induced Liver Fibrosis

A Microfluidic Device for Long‐Term Maintenance of Organotypic Liver Cultures

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