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Updated: Jun 24, 2026
Murine Precision-Cut Liver Slices as an Ex Vivo Model of Liver Biology
Published on: March 14, 2020
Effects of TNF-α in Human Precision-Cut Liver Slices and its Implications for Metabolic Dysfunction-Associated
Ke Luo1, Mei Li1, Alan R Gorter1
1Department of Pharmaceutical Technology and Biopharmacy, Groningen Research Institute of Pharmacy, University of Groningen, Deusinglaan 1, Groningen, 9713 AV, the Netherlands.
Abstract:
Metabolic dysfunction-associated steatohepatitis (MASH) represents a severe form of metabolic dysfunction-associated steatotic liver disease (MASLD), largely due to metabolic dysregulation and sustained liver inflammation. TNF-α plays a pivotal role in MASH pathogenesis by inducing cell death, inflammation, and decreased insulin sensitivity. In this study we investigated the effects of TNF-α in human precision-cut liver slices (PCLS) under healthy or steatotic conditions, to provide insights into MASH pathogenesis. PCLS were prepared from human liver tissue and cultured in control (WEGG) and hyper-nutritive (GFIPO) mediums with or without TNF-α (50 ng/mL) for 96 h. Viability was assessed via ATP content, lipid accumulation by triglyceride (TG) assay, and transcriptomic changes through Next-Generation Sequencing. The protein levels of cytokines, chemokines, and fibrotic mediators released from PCLS were quantified using Luminex assay and ELISA. TNF-α significantly altered the transcriptional profiles in PCLS, inducing pro-inflammatory and pro-fibrotic signaling, and downregulating lipid metabolic processes in both WEGG and GFIPO media. TNF-α showed a trend in elevating intracellular TG in both conditions, albeit not statistically significant. On protein levels, TNF-α supplementation to WEGG medium induced the expression of IL8, CCL2, CCL19, PDGF-AB/BB, TGF-α, and MMP9. GFIPO medium alone induced inflammatory and fibrotic responses indicated by elevated levels of IL8, CCL2, and Pro-collagen 1A1. GFIPO medium with TNF-α supplementation further exacerbated the inflammatory and fibrotic responses, characterized by increased release of cytokines and growth factors. This human PCLS model effectively demonstrated the co-occurrence of key features of MASH, such as steatosis, inflammation, and fibrosis, highlighting the impact of metabolic stress and inflammatory cytokine TNF-α on these disease characteristics, and the potential of the PCLS model in exploring mechanism of MASH progression.
Insights
Tumor necrosis factor-alpha (TNF-α) exacerbates inflammation and fibrosis in metabolic dysfunction-associated steatohepatitis (MASH). Human liver models show TNF-α worsens MASH features, offering insights into disease progression.
Area of Science:
- Hepatology
- Immunology
- Molecular Biology
Background:
- Metabolic dysfunction-associated steatohepatitis (MASH) is a severe liver disease driven by metabolic dysregulation and inflammation.
- Tumor necrosis factor-alpha (TNF-α) is a key mediator in MASH pathogenesis, promoting cell death, inflammation, and insulin resistance.
Purpose of the Study:
- To investigate the impact of TNF-α on human precision-cut liver slices (PCLS) under healthy and steatotic conditions.
- To elucidate the role of TNF-α in MASH pathogenesis using a human PCLS model.
Main Methods:
- Human PCLS were cultured in control (WEGG) or steatotic (GFIPO) media with or without TNF-α.
- Assessed viability (ATP), lipid accumulation (triglycerides), and gene expression (RNA-Seq).
- Quantified protein levels of cytokines, chemokines, and fibrotic mediators (Luminex, ELISA).
Main Results:
- TNF-α altered transcriptional profiles, inducing pro-inflammatory and pro-fibrotic signaling while downregulating lipid metabolism.
- TNF-α increased inflammatory markers (IL8, CCL2) and fibrotic mediators (PDGF, TGF-α, MMP9) in PCLS.
- Combined steatotic conditions and TNF-α exacerbated inflammation and fibrosis.
Conclusions:
- Human PCLS model recapitulates key MASH features: steatosis, inflammation, and fibrosis.
- Metabolic stress and TNF-α significantly impact MASH characteristics.
- PCLS model is valuable for studying MASH progression mechanisms.
