GENE EXPRESSION IN HUMAN HEPATOCYTES IN SUSPENSION AFTER ISOLATION IS SIMILAR TO THE LIVER OF ORIGIN, IS NOT AFFECTED BY HEPATOCYTE COLD STORAGE AND CRYOPRESERVATION, BUT IS STRONGLY CHANGED AFTER HEPATOCYTE PLATING
Abstract
Isolated primary human hepatocytes are a well accepted system for evaluating pharmacological and toxicological effects in humans. However, questions remain regarding how culturing affects the liver-specific functions of the hepatocytes. In addition, cryopreservation could also potentially affect the differentiation state of the hepatocytes. The first aim of the present study was to compare gene expression in freshly isolated primary hepatocytes to that of the liver of origin and to evaluate the expression changes occurring after cryopreservation/thawing, both when maintained in suspension and after plating. The second aim of the present study was to evaluate gene expression in hepatocytes after cold storage of suspensions up to 24 h compared with freshly isolated hepatocytes in suspension. Our results show that the gene expression in freshly isolated human hepatocytes in suspension after isolation is similar to that of the liver of origin. Furthermore, gene expression in primary human hepatocytes in suspension is not affected by hepatocyte cold storage and cryopreservation. However, the gene expression is profoundly affected in monolayer cultures after plating. Specifically, gene expression changes were observed in cultured relative to suspensions of human hepatocytes that are involved in cellular processes such as phase I/II metabolism, basolateral and canalicular transport systems, fatty acid and lipid metabolism, apoptosis, and proteasomal protein recycling. An oxidative s…
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All CYPs with a reported gene expression >30% at 72 h maintained protein expression >60% at 168 h.
“All CYPs with a reported gene expression >30 % at 72 h maintained protein expression >60 % at 168 h.”
From Mechanistic evaluation of primary human hepatocyte culture using global proteomic analysis reveals a selective dedifferentiation profile (2017) · cites this paper as Richert et al. 2006
At 72 h, the mRNA of CYP1A2, 2A6, 2B6, 2C8, 2E1, 3A4, 4A11, 4F2, 4F3 and 27A1 is reported to drop to below 10% of freshly isolated PHH.
“at 72 h, the mRNA of CYP1A2, 2A6, 2B6, 2C8, 2E1, 3A4, 4A11, 4F2, 4F3 and 27A1 is reported to drop to below 10 % of freshly isolated PHH (Richert et al. 2006).”
From Mechanistic evaluation of primary human hepatocyte culture using global proteomic analysis reveals a selective dedifferentiation profile (2017) · cites this paper as Richert et al. 2006
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- Authors
- Richert, L., Liguori, M. J., Abadie, C., Heyd, B., Mantion, G., Halkic, N., & Waring, J. F.
- Affiliations
- Université de Franche-Comté (FR)
- Venue
- Drug Metabolism and Disposition
- Publisher
- Elsevier BV
- Published
- 2006-05
- Volume
- 34
- Issue
- 5
- Pages
- 870-879
- ISSN
- 0090-9556 (ISSN-L)
- OpenAlex
- W2130093373
- Cited by
- 136
- References
- 42
- Citations by year
- 2026: 2 · 2025: 5 · 2024: 1 · 2023: 5 · 2022: 6 · 2021: 13 · 2020: 7 · 2019: 5 · 2018: 6 · 2017: 5 · 2016: 9 · 2015: 10 · 2014: 4 · 2013: 9 · 2012: 11
- FWCI
- 6.08
- Type
- journal article
- Language
- en
- Open access
- closed
- Locations
- Drug Metabolism and Disposition (publishedVersion)
- PubMed (publishedVersion)
- IRIS (submittedVersion)
- SERVAL (Université de Lausanne) (submittedVersion)
- Topics
- Life Sciences › Pharmacology, Toxicology and Pharmaceutics › Pharmacology › Drug-Induced Hepatotoxicity and Protection
- Health Sciences › Medicine › Hepatology › Liver physiology and pathology
- Health Sciences › Medicine › Surgery › Organ Transplantation Techniques and Outcomes
- Keywords
- primary human hepatocytes, gene expression, hepatocyte cryopreservation, cold storage, monolayer culture, hepatocyte differentiation, phase I and II metabolism, apoptosis, oxidative stress response
- MeSH terms
- Cell Separation, Cells, Cultured, Cytological Techniques, Humans, Liver, RNA, Tissue Preservation, Gene Expression, Cryopreservation, Oligonucleotide Array Sequence Analysis, Hepatocytes, In Vitro Techniques
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