The DOI matches its Crossref record; 50 of the 52 references it cites were found in a registry or an index.
Long-Term Culture and Coculture of Primary Rat and Human Hepatocytes
Open accessAbstract
The liver is the largest internal organ in mammals, serving a wide spectrum of vital functions. Loss of liver function due to drug toxicity or viral infection is a major cause of death in the United States. The development of Bioartificial Liver (BAL) devices and the demand for pharmaceutical and cosmetic toxicity screening require the development of long-term hepatocyte culture techniques. However, primary hepatocytes rapidly lose their cuboidal morphology and liver-specific functions over a few days in culture. Accumulation of stress fibers, loss of metabolic function, and cell death are known phenomena. In recent years, several techniques were developed that can support high levels of liver-specific gene expression, metabolic and synthetic function for several weeks in culture. These include the collagen double-gel configuration, hepatocyte spheroids, coculture with endothelial cells, and micropatterned cocultures with 3T3-J2 fibroblasts. This chapter covers the current status of hepatocyte culture techniques, including: hepatocyte isolation, media formulation, oxygen supply, heterotypic cell-cell interactions, and basic functional assays.
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“However, the limited proliferative capacity of PHs, often associated with reduced viability and progressive loss of hepatocyte-specific functions in vitro, has constrained their broader contribution to health and science [39].”
From The Hepatocyte Expansion Paradox: A Review of In Vitro Challenges and Advances (2026) · cites this paper as [39]
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Cites
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“Drug development is currently estimated at $400 million/drug with the majority spent on preclinical screening (5).”
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“One assumption is that spheroids resemble the organization of liver during regeneration (22, 23).”
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Khetani, S. R., & Bhatia, S. N. (2008) · Nature Biotechnology · doi:10.1038/nbt1361 · Cited by 1,219 · cited 2×
“Cell morphology and gene expression were preserved for 4–6 weeks (28).”
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“Culture in sandwich configuration or spheroids relies on the assembly of 3D-like structures, while coculture with endothelial cells or 3T3-J2 fibroblasts depends on heterotypic interactions (15).”
BackgroundHewitt, N. J., Gómez Lechón, M. J., Houston, J. B., Hallifax, D., Brown, H. S., Maurel, P., Kenna, J. G., Gustavsson, L., Lohmann, C., Skonberg, C., et al. (2007) · Drug Metabolism Reviews · doi:10.1080/03602530601093489 · Cited by 735 · cited 1×
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Bile Salt Export Pump: Drug-Induced Liver Injury and Assessment Approaches
Deng, R. (2018) · Methods in pharmacology and toxicology · doi:10.1007/978-1-4939-7677-5_15 · Cited by 1
Ramakrishnan, V. M. (2015) · ThinkIR: The University of Louisville's Institutional Repository (University of Louisville) · doi:10.18297/etd/2319
Metadata
- Authors
- Shulman, M., & Nahmias, Y.
- Affiliations
- Hebrew University of Jerusalem (IL)
- Corresponding author
- Nahmias, Yaakov
- ORCID
- Nahmias, Yaakov
- Venue
- Methods in Molecular Biology
- Publisher
- Humana Press
- Published
- 2012
- Volume
- 945
- Pages
- 287-302
- ISSN
- 1064-3745 (ISSN-L)
- 1940-6029
- PMC
- PMC3781339
- OpenAlex
- W1850526299
- Cited by
- 117
- References
- 52
- Citations by year
- 2026: 1 · 2025: 5 · 2024: 6 · 2023: 8 · 2022: 9 · 2021: 8 · 2020: 19 · 2019: 9 · 2018: 18 · 2017: 13 · 2016: 6 · 2015: 6 · 2014: 2 · 2013: 7
- FWCI
- 15.92
- Type
- book chapter
- Language
- en
- Open access
- green
- www.ncbi.nlm.nih.gov
- Licence
- Locations
- Methods in molecular biology (publishedVersion)
- PubMed (publishedVersion)
- Europe PMC (PubMed Central) (acceptedVersion)
- PubMed Central (acceptedVersion)
- Funders
- National Institute of Diabetes and Digestive and Kidney Diseases (K01DK080241)
- Topics
- Health Sciences › Medicine › Hepatology › Liver physiology and pathology
- Health Sciences › Medicine › Surgery › Tissue Engineering and Regenerative Medicine
- Physical Sciences › Engineering › Biomedical Engineering › 3D Printing in Biomedical Research
- Keywords
- primary hepatocytes, long-term hepatocyte culture, hepatocyte co-culture, bioartificial liver, hepatocyte spheroids, hepatocyte isolation, culture medium formulation, oxygen supply, hepatocyte death, drug toxicity testing
- MeSH terms
- Albumins, Animals, Cell Separation, Enzyme-Linked Immunosorbent Assay, Humans, Oxygen, Perfusion, Time Factors, Urea, Coculture Techniques, Cell Culture Techniques, Hepatocytes, Rats
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