Review Article

 Human Liver Organoids for Modeling Metabolic Dysfunction–Associated Fatty Liver Disease (MAFLD): Advances, Limitations, and Translational Potential

Ji-hye Lee^1,*, Chang-Whan Yoon^2,3,*, Hyunwoo OH^2,4, Jae Jun Lee^1,#, Won Sohn^2,4,# 

Abstract

Background/Objectives: Human liver organoids have rapidly emerged as advanced experimental platforms for modeling metabolic dysfunction-associated fatty liver disease (MAFLD). Compared with conventional 2D hepatocyte cultures and animal models, organoids preserve human tissue architecture, cellular heterogeneity, metabolic function, and gene expression identity, enabling physiologically relevant modeling of lipid accumulation, mitochondrial dysfunction, inflammation, and fibrosis.

Methods: Recent advances allow induction of fatty liver phenotypes using free fatty acids or metabolic triggers, recapitulating steatosis, altered β-oxidation, triglyceride accumulation, and transcriptional reprogramming observed in human disease. Furthermore, fatty liver organoids are being integrated with multi-omics profiling, CRISPR-based functional studies, and AI-guided drug screening to identify therapeutic targets and evaluate drug responses.

Results: Their translational potential has expanded to fibrosis modeling, immune-organoid co-culture, and patient-derived precision medicine applications.

Conclusions: Collectively, organoid-based MAFLD research provides a next-generation platform for mechanistic discovery, biomarker validation, and development of novel metabolic and anti-fibrotic therapeutics.

Keywords

liver organoids, metabolic dysfunction-associated fatty liver disease, fatty liver, nonalcoholic steatohepatitis

Introduction

Metabolic dysfunction–associated fatty liver disease (MAFLD) represents the most prevalent chronic liver disorder worldwide and is closely linked to obesity, insulin resistance, and cardiovascular disease. Traditional 2D hepatocyte models lack metabolic stability, while animal models fail to fully reproduce human genetic drivers, immune responses, and disease progression. Organoid technology has emerged as a transformative tool by enabling long-term culture of human tissue–like structures capable of recapitulating disease-specific cellular, metabolic, and transcriptional features.

Liver Organoid Technology and Biological Advantages

Organoids preserve hepatocyte polarity, bile duct–like structures, metabolic zoning, and mitochondrial networks, enabling realistic modeling of MAFLD pathobiology.

Modeling MAFLD Pathogenesis in Organoids

Human liver organoids are capable of recapitulating key pathological hallmarks of metabolic dysfunction–associated fatty liver disease (MAFLD) and have become a physiologically relevant model for dissecting disease mechanisms. One of the earliest and most reproducible features modeled in organoids is steatosis, where exposure to free fatty acids such as oleic and palmitic acid leads to triglyceride accumulation, lipid droplet formation, and impaired lipid flux, closely mirroring human hepatic steatosis patterns [1-3].

In addition to lipid overload, organoids exhibit metabolic reprogramming characteristic of MAFLD, including reduced β-oxidation, impaired oxidative phosphorylation, and mitochondrial stress responses associated with altered energy metabolism [4, 5]. Beyond metabolic changes, organoid platforms have enabled modeling of inflammatory signaling, particularly when co-cultured with immune cells. These systems activate pro-inflammatory cytokine pathways, including TNF-α, IL-6, and downstream NF-κB signaling, recapitulating the inflammatory transition from simple steatosis to steatohepatitis [6].

Finally, organoids can be engineered to model fibrosis, either through co-culture with hepatic stellate cells or via TGF-β stimulation. These methods result in extracellular matrix deposition, increased collagen production, and transcriptional activation of pro-fibrogenic programs, reflecting fibrosis progression observed in advanced MAFLD [7-9]. Together, these findings demonstrate that liver organoids provide a scalable and translationally relevant platform for modeling the full spectrum of MAFLD pathogenesis.

Organoid-Based Platforms for Therapeutic Discovery

Human liver organoids are increasingly being applied as next-generation platforms for therapeutic discovery in metabolic dysfunction-associated fatty liver disease (MAFLD). Owing to their ability to maintain human-specific metabolic function, cellular diversity, and disease-associated transcriptional profiles, organoids enable physiologically relevant drug response screening, allowing evaluation of lipid-lowering compounds, mitochondrial modulators, and anti-fibrotic drugs under controlled disease conditions [3, 4].

The figure 1 illustrates how liver tissue derived from mice fed a choline-deficient L-amino acid–defined (CDAA) diet exhibits region-specific pathological changes and distinct organoid-forming capacities. Gross liver morphology demonstrates a transition from normal appearance to severe steatosis and fibrotic remodeling after CDAA feeding, reflecting typical features of diet-induced nonalcoholic steatohepatitis (NASH). To evaluate how these pathological changes influence epithelial stem/progenitor behavior, organoids were established from anatomically defined regions of the liver—the peripheral lobe (B) and central parenchymal area adjacent to the portal triad (C). This supports the concept that spatial heterogeneity within the diseased liver profoundly shapes progenitor cell responses and organoid development. 

Furthermore, organoids are highly compatible with CRISPR-based functional genomics, enabling targeted perturbation of metabolic regulators and fibrosis-associated genes to validate causal pathways and therapeutic targets [10]. The integration of multi-omics and transcriptome-guided profiling has accelerated biomarker discovery, linking drug-induced molecular signatures with treatment response and disease reversal [7].

Recent advances have also introduced AI-assisted drug prioritization frameworks and patient-derived organoid platforms, enabling prediction of individualized therapeutic efficacy and facilitating precision medicine approaches in MAFLD [3, 5, 6, 11, 12]. Collectively, ongoing translational studies are evaluating agents including GLP-1 analogs, free fatty acid synthesis inhibitors, mitochondrial activators, and anti-fibrotic compounds using organoid-based screening systems, highlighting their expanding role in therapeutic development [11, 13-15].

Figure 1. Establishment of mouse liver organoids in a region-dependent manner. A. Gross morphology of mouse livers following CDAA (choline-deficient L-amino acid-defined) diet feeding, showing progressive steatosis and fibrosis. Schematic illustration indicates anatomical sampling regions, including the extrahepatic biliary region, peripheral lobe region (B), and central parenchymal region adjacent to the portal triad (C). (B,C) Brightfield images of organoids derived from each anatomical region showing differences in morphology and growth dynamics during early culture. Corresponding hematoxylin and eosin (H&E) staining demonstrating distinct epithelial organization and lumen formation across samples. Scale bars = 50 µm.

Clinical Translation and Precision Medicine

Patient-derived liver organoids offer a unique platform for translational research because they retain patient-specific metabolic features, genomic signatures, and disease phenotypes that are often lost in conventional 2D hepatocyte cultures or animal models [16, 17]. These models enable personalized drug response profiling, allowing researchers to evaluate inter-individual variability in therapeutic efficacy, toxicity, and metabolic adaptation, which is particularly relevant in MAFLD where disease trajectories and treatment outcomes vary widely between patients [1, 14, 17, 18].

Additionally, organoids facilitate biomarker discovery, as multi-omics profiling—including transcriptomics, proteomics, and metabolomics—can be integrated to identify molecular indicators predictive of fibrosis progression or treatment response [19]. Combined with CRISPR and lineage tracing approaches, organoids also enable predictive modeling of disease progression, including the transition from steatosis to fibrosis, cirrhosis, or hepatocellular carcinoma (HCC) [10].

With the emergence of high-throughput drug testing, AI-based therapeutic prediction, and biobanking of patient-derived organoids, these models serve as a functional bridge between preclinical experimentation and individualized therapy development. As such, liver organoids are positioned to accelerate precision medicine by enabling patient-tailored treatment decisions and stratified therapeutic strategies in metabolic liver disease [2, 5, 8, 20].

Limitations and Future Directions

Despite significant progress, current liver organoid platforms face several limitations that impede full clinical translation. A major challenge is the absence of complete physiological components—such as immune cells, endothelial networks, and endocrine signals—which restricts accurate modeling of inflammation, fibrosis, and systemic metabolic regulation seen in MAFLD [21]. Additionally, variability in culture protocols, extracellular matrix sources, and differentiation efficiencies results in batch-to-batch inconsistency, limiting reproducibility across laboratories [22]. Scalability also remains a barrier, particularly for high-throughput screening and therapeutic manufacturing, as most organoid platforms rely on labor-intensive manual handling and costly reagents [23].

To overcome these limitations, emerging engineering strategies are being explored, including vascularized organoids, multi-lineage immune–organoid coculture, and microfluidic liver-on-chip systems, which aim to recapitulate physiological perfusion, immune-tissue interactions, and multi-organ crosstalk [24]. Integration of automation, biofabrication, and AI-driven quality control may further enhance reproducibility and scalability, advancing organoid systems toward clinical-grade applications. As these innovations converge, organoids are anticipated to evolve from research tools into platforms supporting biomarker validation, therapeutic testing, and future regenerative medicine strategies [25, 26].

Discussions

Human liver organoids provide a disease-relevant platform for studying metabolic dysfunction-associated fatty liver disease (MAFLD), capturing key pathological features such as steatosis, mitochondrial dysfunction, inflammation, and fibrosis. Unlike conventional 2D hepatocyte systems, organoids retain patient-specific genetic and metabolic signatures, enabling mechanistic studies and modeling of disease heterogeneity. Their integration with technologies such as CRISPR screening, multi-omics profiling, and AI-assisted drug prediction has advanced target discovery and therapeutic evaluation. Patient-derived organoids further support precision medicine, facilitating individualized drug response testing and biomarker development.

However, several limitations remain, including the lack of immune, vascular, and endocrine components, culture variability across laboratories, and challenges in large-scale production. Advancements such as vascularized and immune-integrated organoids, organ-on-chip systems, and automated bioprocessing may improve physiological relevance and scalability. As these innovations mature, liver organoids are positioned to accelerate translational research and support the development of personalized therapies for MAFLD.

Conclusions

Human liver organoids have emerged as a transformative platform for modeling metabolic dysfunction-associated fatty liver disease (MAFLD), providing biologically relevant systems that retain patient-specific genomic, metabolic, and structural features. By enabling mechanistic interrogation of steatosis, inflammation, fibrosis progression, and tumorigenic transition, organoid models bridge a critical gap between traditional in vitro hepatocyte systems and in vivo animal models. Their expanding integration with multi-omics profiling, CRISPR gene editing, immune and vascular coculture systems, and AI-guided therapeutic prediction is accelerating biomarker discovery and target validation.

As patient-derived liver organoids increasingly support personalized drug response profiling and therapeutic stratification, they hold strong potential to reshape precision medicine strategies for MAFLD and downstream complications including fibrosis and hepatocellular carcinoma. Nevertheless, challenges related to scalability, standardization, vascularization, and immune integration remain barriers to full clinical deployment. Continued advances in bioengineering, microfluidics, and organ-on-chip technologies are expected to enhance physiological fidelity and translational applicability.

Collectively, the convergence of organoid engineering and precision therapeutics positions liver organoids as a cornerstone for next-generation MAFLD research, with the capacity to accelerate the development of metabolic, anti-fibrotic, and immunomodulatory therapies toward clinical implementation.

Conflict of Interest

The authors have no conflicts of interest to declare and agreed to the published version of the manuscript

Author Contributions

The contributions of each author to this study are summarized as follows. JHL, CWY, HWO, JKL and WS conceptualized and designed the overall research framework. Data curation was conducted by JHL, CWY and JJL, while HWO and WS were responsible for formal data analysis. Funding for the study was acquired by JJL and WS. The experimental investigation was carried out by JJL and WS. Methodological design and refinement were performed collaboratively by JHL, CWY and HWO. Validation and visualization of the results were undertaken by JHL, CWY, HWO, and WS. The original draft of the manuscript was written by JHL, CWY, and JJL, and the final version of the manuscript was reviewed and edited by JHL, CWY, and WS. All authors approved the final version.

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