| Title: | How to establish a novel liver cell culture system that resembles the in vivo liver microenvironment |
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| Authors: | ID Madorran, Eneko (Author) |
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| Files: |  RAZ_Madorran_Eneko_2025.pdf (1,11 MB)
MD5: 0C2EA9917C541AE473FE655184C1B01A
 https://doi.org/10.1002/cpz1.70172
https://currentprotocols.onlinelibrary.wiley.com/doi/10.1002/cpz1.70172
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| Language: | English |
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| Work type: | Scientific work |
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| Typology: | 1.01 - Original Scientific Article |
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| Organization: | MF - Faculty of Medicine
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| Abstract: | Subtle physiological patterns within the human organism, such as molecular fluctuations or systemic adaptations, often remain hidden from direct observation due to the inherent variability and noise within biological samples. The liver, a vital organ essential for systemic regulation and toxicological assessment, presents this challenge due to its heightened activity, which can influence enzyme dynamics and metabolic processes. Unlike direct observation, in vitro liver models offer a more precise means of understanding these trends, providing insights into the organ's (patho)physiology, and serving as a platform for toxicity evaluation. However, current liver models lack essential features required to faithfully replicate the liver's microenvironment, resulting in reduced accuracy in toxicity assessments. Furthermore, while researchers emphasize mechanistic insights, such as the molecular processes governing glucose metabolism and cellular energy production, clinicians focus on tangible health outcomes, like blood glucose levels in patients. This disconnect between the objectives and methodologies of basic researchers and clinicians amplifies this gap, hindering effective translational research. Addressing these challenges, a novel liver cell culture system that resembles the in vivo liver microenvironment with clinical instrumentation has been proposed to enhance current liver models, improving their capacity to emulate in vivo conditions. This study introduces a novel liver cell culture system, utilizing a 96-well plate format incorporating hepatic sinusoidal endothelial cells, hepatic stellate cells, Kupffer cells, and hepatocytes to replicate the liver microenvironment. The model integrates clinical instrumentation, specifically a biochemical analyzer to ensure biomarkers closely align with those observed in clinical diagnostics. This design enables researchers to fine-tune conditions that mimic the liver's microanatomy and physiological responses, enhancing its translational potential for toxicity assessments. The approach involves primary cell culture preparation, supernatant analysis through a clinical biochemistry analyzer, and cell viability assessment using the Membrane Potential Cell Viability Assay (MPCVA) method. Additionally, advanced imaging techniques and data analysis tools are incorporated to refine the model's capabilities and ensure greater translatability to clinical applications. |
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| Keywords: | in vitro toxicity, liver (patho)physiology, liver in vitro model, membrane potential cell viability assay, translational research |
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| Publication status: | Published |
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| Publication version: | Version of Record |
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| Publication date: | 12.07.2025 |
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| Publisher: | Wiley Periodicals LLC |
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| Year of publishing: | 2025 |
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| Number of pages: | Str. 1-14 |
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| Numbering: | Letn. 5, Št. 7 |
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| PID: | 20.500.12556/DKUM-94079  |
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| UDC: | 60 |
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| ISSN on article: | 2691-1299 |
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| COBISS.SI-ID: | 244120067 |
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| DOI: | 10.1002/cpz1.70172 |
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| Publication date in DKUM: | 14.08.2025 |
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| Views: | 0 |
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| Downloads: | 0 |
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| Metadata: |    |
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| Categories: | Misc.
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