Abstract

Molecular and cellular research modalities for the study of liver pathologies have been tremendously improved over the recent decades. Advanced technologies offer novel opportunities to establish cell isolation techniques with excellent purity, paving the path for 2D and 3D microscopy and high-throughput assays (e.g., bulk or single-cell RNA sequencing). The use of stem cell and organoid research will help to decipher the pathophysiology of liver diseases and the interaction between various parenchymal and non-parenchymal liver cells. Furthermore, sophisticated animal models of liver disease allow for the in vivo assessment of fibrogenesis, portal hypertension and hepatocellular carcinoma (HCC) and for the preclinical testing of therapeutic strategies. The purpose of this review is to portray in detail novel in vitro and in vivo methods for the study of liver cell biology that had been presented at the workshop of the 8th meeting of the European Club for Liver Cell Biology (ECLCB-8) in October of 2018 in Bonn, Germany.

Highlights

  • Cell culture techniques are important tools for the study of pathogenesis and treatment of liver diseases

  • Two principal populations can be differentiated by their ontogeny, polarization and function during injury and resolution [71], namely Kupffer cells (KCs) and monocytes, and their functional behavior in liver diseases could become a target for novel therapeutics (Table 1) [38]

  • In order to study the mechanisms produced in non-alcoholic steatohepatitis (NASH) pathology and the potential therapeutic targets, the models most commonly used are as follows [42]: dietary models, such as HFD, high-fructose diet [43], cholesterol and cholate diet [37], MCD diet [95] and choline-deficient l-amino acid-defined (CDAA) diet [96]; genetic models, some of which act by promoting fat synthesis, while others act by inhibiting lipid peroxidation [97] and impeding fat transport (ApoE knock-out mice) [35]; and chemical models, such as CCL4 [98], tetracycline [99] and streptozotocin in combination with HFD [100]

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Summary

Introduction

Cell culture techniques are important tools for the study of pathogenesis and treatment of liver diseases. The use of 2D microscopy, 3D microscopy, intravital microscopy, flow cytometry and cell isolation for subsequent functional experiments or expression analyses is expected to significantly increase characterization of hepatic cell populations and their interactions with circulating immune cells. These methods have been used to study the impact of hepatic macrophages and monocytes on steatosis, inflammation, hepatocellular injury, HSCs activation and angiogenesis [4]. The purpose of this review is to present technological tools that will enhance our ability to isolate and purify the different cell populations and other strategies to study the full spectrum of liver disease (Figure 1)

HSCs Isolation and Immortalized HSCs Lines
Analysis of Liver Cells from Different Human Donors and Co-Culture Models
Isolation and Characterization of Liver Macrophages
Cellular Models in Biliary Research
Angiogenesis and Gene Expression Inhibition
The Leading Models of Experimental ALD
Mouse Models for Non-Alcoholic Fatty Liver Disease
Mouse Models for Hepatocellular Carcinoma
In Vivo Setup of Liver Disease Models for Surgery and Hemodynamic Assessments
Conclusion
Findings
32. Liquid Diet Technique of ethanol administration
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