Viability, function and morphological integrity of precision-cut liver slices during prolonged incubation: Effects of culture medium

Starokozhko, Viktoriia; Abza, Getahun Befirdu; Maessen, Hedy C; Merema, Marjolijn T.; Kuper, Frieke; Groothuis, Geny M. M.

doi:10.1016/j.tiv.2015.10.008

Cited by 23 publications

(24 citation statements)

References 73 publications

(87 reference statements)

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“…Protein content of the hPCLS was measured according to Lowry by using the Bio-Rad DC Protein Assay (Bio-Rad, Munich, Germany) using a bovine serum albumin calibration curve (Lowry et al 1951) as previously described (Starokozhko et al 2015). …”

Section: Methodsmentioning

confidence: 99%

“…The glycogen content of hPCLS was determined by the periodic acid-Schiff (PAS) staining as described previously by Schaart et al (2004), with some modifications as described before (Starokozhko et al 2015). Staining for fibrillary collagen was performed on 4-µm paraffin sections using picrosirius red (Sigma, Gillingham, UK).…”

Section: Methodsmentioning

confidence: 99%

“…Improved viability and functionality of the slices can be achieved by improving culture conditions such as medium composition (Olinga et al 1997; Starokozhko et al 2015). For example, a recent study on rat PCLS showed that the medium composition has a large impact on tissue viability and functions following 5 days of incubation (Starokozhko et al 2015).…”

Section: Introductionmentioning

confidence: 99%

“…For example, a recent study on rat PCLS showed that the medium composition has a large impact on tissue viability and functions following 5 days of incubation (Starokozhko et al 2015). It is generally known that for a proper prediction of drug disposition and toxicity, it is very important to use human cells or tissues because of large species differences in these functions (Hadi et al 2013).…”

Section: Introductionmentioning

confidence: 99%

“…Williams’ Medium E (WME) was chosen as a standard cell culture medium that is commonly used for PCLS incubation (Duryee et al 2014; Jetten et al 2014; Westra et al 2014). As a second medium we chose RegeneMed ® , which was designed and used for long-term culture of primary human liver cells (Kostadinova et al 2013) and which we tested on rat PCLS before (Starokozhko et al 2015). As a third medium, we tested Cellartis ® Hepatocyte Maintenance Medium (Takara Bio Europe AB), which was originally designed as maintenance medium for induced pluripotent stem-cell-derived hepatocytes, to maintain viability, differentiation, and liver functions.…”

Section: Introductionmentioning

confidence: 99%

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Maintenance of drug metabolism and transport functions in human precision-cut liver slices during prolonged incubation for 5 days

et al. 2016

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Human precision-cut liver slices (hPCLS) are a valuable ex vivo model that can be used in acute toxicity studies. However, a rapid decline in metabolic enzyme activity limits their use in studies that require a prolonged xenobiotic exposure. The aim of the study was to extend the viability and function of hPCLS to 5 days of incubation. hPCLS were incubated in two media developed for long-term culture of hepatocytes, RegeneMed®, and Cellartis®, and in the standard medium WME. Maintenance of phase I and II metabolism was studied both on gene expression as well as functional level using a mixture of CYP isoform-specific substrates. Albumin synthesis, morphological integrity, and glycogen storage was assessed, and gene expression was studied by transcriptomic analysis using microarrays with a focus on genes involved in drug metabolism, transport and toxicity. The data show that hPCLS retain their viability and functionality during 5 days of incubation in Cellartis® medium. Albumin synthesis as well as the activity and gene expression of phase I and II metabolic enzymes did not decline during 120-h incubation in Cellartis® medium, with CYP2C9 activity as the only exception. Glycogen storage and morphological integrity were maintained. Moreover, gene expression changes in hPCLS during incubation were limited and mostly related to cytoskeleton remodeling, fibrosis, and moderate oxidative stress. The expression of genes involved in drug transport, which is an important factor in determining the intracellular xenobiotic exposure, was also unchanged. Therefore, we conclude that hPCLS cultured in Cellartis® medium are a valuable human ex vivo model for toxicological and pharmacological studies that require prolonged xenobiotic exposure.Electronic supplementary materialThe online version of this article (doi:10.1007/s00204-016-1865-x) contains supplementary material, which is available to authorized users.

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Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Maintenance of drug metabolism and transport functions in human precision-cut liver slices during prolonged incubation for 5 days

et al. 2016

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Time‐Resolved Quantification of Nanoparticle Uptake, Distribution, and Impact in Precision‐Cut Liver Slices

Bartucci

Åberg

Melgert

et al. 2020

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Much effort within the nanosafety field is currently focused on the use of advanced in vitro models to reduce the gap between in vitro and in vivo studies. Within this context, precision‐cut tissue slices are a unique ex vivo model to investigate nanoparticle impact using live tissue from laboratory animals and even humans. However, several aspects of the basic mechanisms of nanoparticle interactions with tissue have not yet been elucidated. To this end, liver slices are exposed to carboxylated and amino‐modified polystyrene known to have a different impact on cells. As observed in standard cell cultures, amino‐modified polystyrene nanoparticles induce apoptosis, and their impact is affected by the corona forming on their surface in biological fluids. Subsequently, a detailed time‐resolved study of nanoparticle uptake and distribution in the tissue is performed, combining fluorescence imaging and flow cytometry on cells recovered after tissue digestion. As observed in vivo, the Kupffer cells accumulate high nanoparticle amounts and, interestingly, they move within the tissue towards the slice borders. Similar observations are reproduced in liver slices from human tissue. Thus, tissue slices can be used to reproduce ex vivo important features of nanoparticle outcomes in the liver and study nanoparticle impact on real tissue.

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Challenges on the road to a multicellular bioartificial liver

Starokozhko

Groothuis

2017

J Tissue Eng Regen Med

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Over recent years, the progress in the development of a bioartificial liver (BAL) as an extracorporeal device or as a tissue engineered transplantable organ has been immense. However, many important BAL characteristics that are necessary to meet clinical demands have not been sufficiently addressed. This review describes the existing challenges in the development of a BAL for clinical applications, highlighting multicellularity and sinusoidal microarchitecture as crucial BAL characteristics to fulfil various liver functions. Currently available sources of nonparenchymal liver cells, such as endothelial cells, cholangiocytes and macrophages, used in BAL development are defined. Also, we discuss the recent studies on the reconstruction of the complex liver sinusoid microarchitecture using various liver cell types. Moreover, we highlight some other aspects of a BAL, such as liver zonation and formation of a vascular as well as biliary network for an adequate delivery, biotransformation and removal of substrates and waste products. Finally, the benefits of a multicellular BAL for the pharmaceutical industry are briefly described.

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Viability, function and morphological integrity of precision-cut liver slices during prolonged incubation: Effects of culture medium

Cited by 23 publications

References 73 publications

Maintenance of drug metabolism and transport functions in human precision-cut liver slices during prolonged incubation for 5 days

Maintenance of drug metabolism and transport functions in human precision-cut liver slices during prolonged incubation for 5 days

Time‐Resolved Quantification of Nanoparticle Uptake, Distribution, and Impact in Precision‐Cut Liver Slices

Challenges on the road to a multicellular bioartificial liver

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