ArticleMolecular pharmaceutics2010
Influence of seeding density and extracellular matrix on bile Acid transport and mrp4 expression in sandwich-cultured mouse hepatocytes.
Article in Molecular pharmaceutics, 2010. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 17 papers.
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Who cites it
17 citing papers in PubMed, 43 citations in OpenAlex.
- Activation of cAMP (EPAC2) signaling pathway promotes hepatocyte attachment.Scientific reports · 2023Article
- In Vitro Assay System to Detect Drug-Induced Bile Acid-Dependent Cytotoxicity Using Hepatocytes.Methods in molecular biology (Clifton, N.J.) · 2022Article
- Prominin-1-Radixin axis controls hepatic gluconeogenesis by regulating PKA activity.EMBO reports · 2020Article
- The importance of transporters and cell polarization for the evaluation of human stem cell-derived hepatic cells.PloS one · 2020Article
- Current progress in hepatic tissue regeneration by tissue engineering.Journal of translational medicine · 2019Review
- Evaluation of Drug Biliary Excretion Using Sandwich-Cultured Human Hepatocytes.European journal of drug metabolism and pharmacokinetics · 2019Review
- Prolongation of liver-specific function for primary hepatocytes maintenance in 3D printed architectures.Organogenesis · 2018Article
- Decellularization and Solubilization of Porcine Liver for Use as a Substrate for Porcine Hepatocyte Culture: Method Optimization and Comparison.Cell transplantation · 2017Article
- Three-dimensional (3D) printing of mouse primary hepatocytes to generate 3D hepatic structure.Annals of surgical treatment and research · 2017Article
- Activation of the constitutive androstane receptor inhibits gluconeogenesis without affecting lipogenesis or fatty acid synthesis in human hepatocytes.Toxicology and applied pharmacology · 2014Article
- Co-regulation of primary mouse hepatocyte viability and function by oxygen and matrix.Biotechnology and bioengineering · 2014Article
- Human placental lactogen induces CYP2E1 expression via PI 3-kinase pathway in female human hepatocytes.Drug metabolism and disposition: the biological fate of chemicals · 2014Article
- Article
- Successful mouse hepatocyte culture with sandwich collagen gel formation.Journal of the Korean Surgical Society · 2013Article
- Comparison of culture media for bile Acid transport studies in primary human hepatocytes.Journal of clinical and experimental hepatology · 2012Article
- Evaluation of an in vitro toxicogenetic mouse model for hepatotoxicity.Toxicology and applied pharmacology · 2010Article
- Sandwich-cultured hepatocytes: an in vitro model to evaluate hepatobiliary transporter-based drug interactions and hepatotoxicity.Drug metabolism reviews · 2010Review
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2 authors at 1 institution in 1 country.
Funding
Abstract
This study was undertaken to examine the influence of seeding density, extracellular matrix and days in culture on bile acid transport proteins and hepatobiliary disposition of the model bile acid taurocholate. Mouse hepatocytes were cultured in a sandwich configuration on six-well Biocoat plates with an overlay of Matrigel (BC/MG) or gelled-collagen (BC/GC) for 3 or 4 days at seeding densities of 1.0, 1.25, or 1.5 x 10(6) cells/well. The lower seeding densities of 1.0 and 1.25 x 10(6) cells/well resulted in good hepatocyte morphology and bile canalicular network formation, as visualized by 5-(and 6)-carboxy-2',7'-dichlorofluorescein accumulation. In general, taurocholate cellular accumulation tended to increase as a function of seeding density in BC/GC; cellular accumulation was significantly increased in hepatocytes cultured in BC/MG compared to BC/GC at the same seeding density on both days 3 and 4 of culture. In general, in vitro intrinsic biliary clearance of taurocholate was increased at higher seeding densities. Levels of bile acid transport proteins on days 3 and 4 were not markedly influenced by seeding density or extracellular matrix except for multidrug resistance protein 4 (Mrp4), which was inversely related to seeding density. Mrp4 levels decreased approximately 2- to 3-fold between seeding densities of 1.0 x 10(6) and 1.25 x 10(6) cells/well regardless of extracellular matrix; an additional approximately 3- to 5-fold decrease in Mrp4 protein was noted in BC/GC between seeding densities of 1.25 x 10(6) and 1.5 x 10(6) cells/well. Results suggest that seeding density, extracellular matrix and days in culture profoundly influence Mrp4 expression in sandwich-cultured mouse hepatocytes. Primary mouse hepatocytes seeded in a BC/MG configuration at densities of 1.25 x 10(6) cells/well and 1.0 x 10(6), and cultured for 3 days, yielded optimal transport based on the probes studied. This work demonstrates the applicability of the sandwich-cultured model to mouse hepatocytes.
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