ArticleExperimental biology and medicine (Maywood, N.J.)2015
Featured Article: Isolation, characterization, and cultivation of human hepatocytes and non-parenchymal liver cells.
Article in Experimental biology and medicine (Maywood, N.J.), 2015. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 54 papers.
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Who cites it
54 citing papers in PubMed, 102 citations in OpenAlex.
- Semi-automated isolation of parenchymal and non-parenchymal liver cells from mice and humans with enhanced stellate cell fraction.Cell & bioscience · 2026Article
- Hepatic Arterial Flow-Induced Portal Tract Fibrosis in Portal Hypertension: The Role of VCAM-1 and Osteopontin-Expressing Macrophages.Cellular and molecular gastroenterology and hepatology · 2026Article
- Convergent Metabolic Pathways in MASH Therapeutics: An AMPK-Centric Analysis.Journal of cellular and molecular medicine · 2026Review
- Ex vivo Drug Metabolism Using Primary Human Hepatocytes for the Detection of Metabolite-Dependent Antibodies to Red Blood Cells.Transfusion medicine and hemotherapy : offizielles Organ der Deutschen Gesellschaft fur Transfusionsmedizin und Immunhamatologie · 2025Article
- Expression of Selected Pharmacologically Relevant Transporters in Murine Non-Parenchymal Liver Cells Compared to Hepatocytes.International journal of molecular sciences · 2025Article
- High-yield isolation of primary human hepatocytes from small liver samples.In vitro models · 2025Article
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- Impact of Hedgehog modulators on signaling pathways in primary murine and human hepatocytes in vitro: insights into liver metabolism.Archives of toxicology · 2025Article
- Generation of Multicellular 3D Liver Organoids From Induced Pluripotent Stem Cells as a Tool for Modelling Liver Diseases.Bio-protocol · 2024Article
- Sex hormones differently regulate lipid metabolism genes in primary human hepatocytes.BMC endocrine disorders · 2024Article
- Cell atlas of the regenerating human liver after portal vein embolization.Nature communications · 2024Article
- High-Voltage Electrostatic Field Hydrogel Microsphere 3D Culture System Improves Viability and Liver-like Properties of HepG2 Cells.International journal of molecular sciences · 2024Article
- The Sympathetic Nervous System Promotes Hepatic Lymphangiogenesis, which Is Protective Against Liver Fibrosis.The American journal of pathology · 2023Article
- Bioengineering liver microtissues for modeling non-alcoholic fatty liver disease.EXCLI journal · 2023Review
- Optimization of extracellular matrix for primary human hepatocyte cultures using mixed collagen-Matrigel matrices.EXCLI journal · 2023Article
- Multiparametric Flow Cytometry-Based Immunophenotyping of Mouse Liver Immune Cells.Methods and protocols · 2022Article
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- Advancements in the Alcohol-Associated Liver Disease Model.Biomolecules · 2022Review
- Enrichment Methods for Murine Liver Non-Parenchymal Cells Differentially Affect Their Immunophenotype and Responsiveness towards Stimulation.International journal of molecular sciences · 2022Article
Corrections and comments
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Authors and funding
7 authors at 3 institutions in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Primary human hepatocytes (PHH) are considered to be the gold standard for in vitro testing of xenobiotic metabolism and hepatotoxicity. However, PHH cultivation in 2D mono-cultures leads to dedifferentiation and a loss of function. It is well known that hepatic non-parenchymal cells (NPC), such as Kupffer cells (KC), liver endothelial cells (LEC), and hepatic stellate cells (HSC), play a central role in the maintenance of PHH functions. The aims of the present study were to establish a protocol for the simultaneous isolation of human PHH and NPC from the same tissue specimen and to test their suitability for in vitro co-culture. Human PHH and NPC were isolated from tissue obtained by partial liver resection by a two-step EDTA/collagenase perfusion technique. The obtained cell fractions were purified by Percoll density gradient centrifugation. KC, LEC, and HSC contained in the NPC fraction were separated using specific adherence properties and magnetic activated cell sorting (MACS®). Identified NPC revealed a yield of 1.9 × 10(6) KC, 2.7 × 10(5) LEC and 4.7 × 10(5) HSC per gram liver tissue, showing viabilities >90%. Characterization of these NPC showed that all populations went through an activation process, which influenced the cell fate. The activation of KC strongly depended on the tissue quality and donor anamnesis. KC became activated in culture in association with a loss of viability within 4-5 days. LEC lost specific features during culture, while HSC went through a transformation process into myofibroblasts. The testing of different culture conditions for HSC demonstrated that they can attenuate, but not prevent dedifferentiation in vitro. In conclusion, the method described allows the isolation and separation of PHH and NPC in high quality and quantity from the same donor.
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Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.