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Hepatocytes in collagen sandwich: evidence for transcriptional and translational regulation.
Open accessAbstract
The influence of extracellular matrix configuration on the tissue-specific function of cultured hepatocytes was investigated. Adult rat hepatocytes sandwiched between two layers of collagen gel were compared to cells cultured on a single layer of collagen gel for differences in the total RNA content, the level of albumin-specific mRNA, the rate of albumin gene transcription, and the rate of albumin mRNA translation. Adult hepatocytes in the sandwich system maintained the level of albumin mRNA similar to that found in the normal liver for at least six weeks, whereas the level of albumin mRNA declined rapidly in the single gel system. After one week of culture, hepatocytes in the single gel system could be induced to recover the high level of albumin mRNA and albumin production when a second layer of collagen gel was overlaid at that time. Furthermore, sandwiched hepatocytes maintained significantly higher transcriptional activity compared to cells in the single gel system. In addition to transcriptional control, the ultimate rate of albumin production was shown to depend on the rate of translation, which increased with culture time and reached a plateau in one to two weeks. This increase in translational activity over time in culture was observed in both the sandwich and the single gel systems and, thus, appeared to be independent of the configuration of extracellular matrix.
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“The sandwich culture method, e.g., placing cells between two layers comprised of ECM, polyacrylamide, or collagen, has long been demonstrated to produce hepatocyte cell cultures, where morphology and function more accurately reproduce in vivo behavior (9,29,34,65,73).”
From Modeling physiological events in 2D vs. 3D cell culture (2017) · cites this paper as [29]
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“Their results showed that the sandwich culture could maintain a proper albumin production rate for at least 6 wk, whereas the one-gel system already had rates that were tenfold lower than normal by the end of the first week (29).”
From Modeling physiological events in 2D vs. 3D cell culture (2017) · cites this paper as [29]
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“The effect of cell polarization in 2D cell culture can be mitigated using a sandwich culture method that adds a layer of ECM atop the cells (9,29,34,65,73) to eliminate apical-basal polarity and provide a mimic of 3D ECM.”
From Modeling physiological events in 2D vs. 3D cell culture (2017) · cites this paper as [29]
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“In 2D cultures, there are decreases in transcriptional genes and rapid changes in morphology (29,34).”
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“Hepatocytes in the liver are surrounded by a complex ECM, and several researchers have shown that conventional 2D culture methods, flasks with or without ECM protein coatings, do not establish well enough the complex microenvironment that hepatocytes need to function (29,34,73).”
From Modeling physiological events in 2D vs. 3D cell culture (2017) · cites this paper as [29]
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Metadata
- Authors
- Dunn, J. C., Tompkins, R. G., & Yarmush, M. L.
- Affiliations
- Harvard University (US)
- Venue
- The Journal of cell biology
- Publisher
- Rockefeller University Press
- Published
- 1992-02-15
- Volume
- 116
- Issue
- 4
- Pages
- 1043-1053
- ISSN
- 0021-9525 (ISSN-L)
- 1540-8140
- PMC
- PMC2289333
- OpenAlex
- W2067188612
- Cited by
- 295
- References
- 52
- Citations by year
- 2026: 2 · 2025: 6 · 2024: 6 · 2023: 7 · 2022: 6 · 2021: 5 · 2020: 6 · 2019: 10 · 2018: 9 · 2017: 4 · 2016: 8 · 2015: 14 · 2014: 8 · 2013: 6 · 2012: 3
- FWCI
- 8.43
- Type
- journal article
- Language
- en
- Open access
- green
- www.ncbi.nlm.nih.gov
- Licence
- cc-by-nc-sa
- Locations
- The Journal of Cell Biology (publishedVersion)
- PubMed Central (publishedVersion, cc-by-nc-sa)
- PubMed (publishedVersion)
- Funders
- National Institutes of Health (5r01dk041709-05, 5k08dk001746-05)
- National Institute of Diabetes and Digestive and Kidney Diseases (DK-01746, R01 DK-41709)
- Topics
- Health Sciences › Medicine › Hepatology › Liver physiology and pathology
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Cell Biology › Cellular Mechanics and Interactions
- Health Sciences › Medicine › Surgery › Tissue Engineering and Regenerative Medicine
- Keywords
- hepatocytes, albumin synthesis, tissue-specific gene expression, hepatocyte culture
- MeSH terms
- Albumins, Animals, Cells, Cultured, Collagen, Extracellular Matrix, Female, Gene Expression Regulation, Kinetics, Liver, Rats, Inbred Lew, RNA, Messenger, Transcription, Genetic, Protein Biosynthesis, Rats
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