OATP1B1/1B3 activity in plated primary human hepatocytes over time in culture
Open accessAbstract
Primary human hepatocytes are widely used as an in vitro model for evaluation of drug metabolism and transport. However, it has been shown that the gene expression of many drug-metabolizing enzymes and transporters change in culture. The aim of the present study was to evaluate the activity of organic anion-transporting polypeptide 1B1 (OATP1B1) and 1B3 (OATP1B3) in plated primary human hepatocytes over time in culture. The uptake kinetics of the OATP1B1/1B3 substrate [(3)H]-estradiol-17β-d-glucuronide was determined in cells from five donors. An extensive and variable decrease in OATP1B1/1B3 activity and/or increase in passive diffusion was observed over time. Already after 6h in culture, the OATP1B1/1B3 activity was not possible to determine in liver cells from one donor, while after 24h, the uptake activity was not measurable in one additional donor. In the other three, the decrease in CL(int) (V(max)/K(m)) values ranged from 15% to 86% after 24h in culture compared to the values measured at 2h. Visual examination of OATP1B1 protein expression by confocal microscopy showed localization to the plasma membrane as expected, and an extensive decrease in OATP1B1 expression over time in culture supported the decline in activity. The significant reduction in SLCO1B1 and SLCO1B3 gene expression over time determined by RT-PCR also supported the loss of OATP1B1/1B3 activity. In conclusion, plated primary human hepatocytes are useful as an in vitro model for OATP1B1/1B3-mediated uptake studies, but the culture time may substantially change the uptake kinetics.
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The expression of OATP1B1, selectively found in the sinusoidal hepatocyte membrane, declines rapidly in hepatocyte cultures.
“The expression of the drug membrane transporter OATP1B1 that is selectively found in the sinusoidal hepatocyte membrane, declines rapidly in hepatocyte cultures (Ulvestad et al., 2011[41]).”
From Optimization of extracellular matrix for primary human hepatocyte cultures using mixed collagen-matrigel matrices (2023) · cites this paper as Ulvestad et al., 2011[41]
The Summary of Papers reports an extensive decrease in OATP1B1 expression over time in plated primary human hepatocytes, and §4.1.1 places OATP1B1 at the basolateral membrane in the human liver. The claim is broader than the evidence: the decline was observed in plated cultures, whereas the Abstract reports preserved OATP1B1 expression for at least nine days in 3D bioreactor cultures. (LLM, conf=0.86)
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Godoy, P., Hewitt, N. J., Albrecht, U., Andersen, M. E., Ansari, N., Bhattacharya, S., Bode, J. G., Bolleyn, J., Borner, C., Böttger, J., et al. (2013) · Archives of Toxicology · doi:10.1007/s00204-013-1078-5 · Cited by 1,280 · see citing works
Seidemann, L., Prinz, S., Scherbel, J.-C., Götz, C., Seehofer, D., & Damm, G. (2023) · IfADo - Leibniz Research Centre for Working Environment and Human Factors, Dortmund · doi:10.17179/excli2022-5459 · Cited by 3 · see citing works
“The expression of the drug membrane transporter OATP1B1 that is selectively found in the sinusoidal hepatocyte membrane, declines rapidly in hepatocyte cultures (Ulvestad et al., 2011).”
- Authors
- Ulvestad, M., Björquist, P., Molden, E., Åsberg, A., & Andersson, T. B.
- Affiliations
- University of Oslo (NO)
- Karolinska Institutet (SE)
- Corresponding author
- Andersson, Tommy B.
- Venue
- Biochemical Pharmacology
- Publisher
- Elsevier BV
- Published
- 2011-11
- Volume
- 82
- Issue
- 9
- Pages
- 1219-1226
- ISSN
- 0006-2952 (ISSN-L)
- OpenAlex
- W2072777053
- Cited by
- 40
- References
- 3
- Citations by year
- 2026: 1 · 2025: 1 · 2023: 1 · 2022: 3 · 2021: 1 · 2020: 1 · 2019: 2 · 2018: 2 · 2017: 2 · 2016: 3 · 2015: 3 · 2014: 5 · 2013: 9 · 2012: 6
- FWCI
- 3.14
- Type
- journal article
- Language
- en
- Open access
- bronze
- doi.org
- Locations
- Biochemical Pharmacology (publishedVersion)
- PubMed (publishedVersion)
- Funders
- Norges Forskningsråd
- Topics
- Health Sciences › Medicine › Oncology › Drug Transport and Resistance Mechanisms
- Life Sciences › Pharmacology, Toxicology and Pharmaceutics › Pharmacology › Pharmacogenetics and Drug Metabolism
- Life Sciences › Pharmacology, Toxicology and Pharmaceutics › Pharmacology › Drug-Induced Hepatotoxicity and Protection
- Keywords
- OATP1B1, OATP1B3, organic anion transporters, primary human hepatocytes, uptake kinetics, passive diffusion, SLCO1B1, SLCO1B3
- MeSH terms
- Solute Carrier Organic Anion Transporter Family Member 1B3, Adult, Aged, Cells, Cultured, Estradiol, Female, Gene Expression Regulation, Genotype, Humans, Male, Middle Aged, Time Factors, Hepatocytes, Organic Anion Transporters, Liver-Specific Organic Anion Transporter 1, Organic Anion Transporters, Sodium-Independent
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