Small hepatocytes in culture develop polarized transporter expression and differentiation

Pfändler, Marguerite-Anne Sidler; Höchli, M.; Inderbitzin, Daniel; Meier, Peter J.; Stieger, Bruno

doi:10.1242/jcs.01279

Cited by 33 publications

(10 citation statements)

References 48 publications

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“…Considering the fact that the upregulation of Mrp3 is often observed in cholestatic conditions, the cyst formation may release the pressure of accumulating bile in closed BC and create a flow directly to the cyst. Pfä ndler et al [2004] also demonstrated that time-dependent expression of individual transporters correlating with the maturation of SHs in growing colonies had some striking similarities with the ontogenesis of the hepatocellular bile-salt and organic-anion transport polarity. We have demonstrated that the SH is a committed hepatic progenitor cell that can further differentiate into an MH [Mitaka et al, 1999;Kon et al, 2006].…”

Section: Discussionmentioning

confidence: 81%

Functional expression of organic anion transporters in hepatic organoids reconstructed by rat small hepatocytes

Oshima

Kon

Ooe

et al. 2007

J of Cellular Biochemistry

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Small hepatocytes (SHs) are hepatic progenitor cells with hepatic characteristics. They can proliferate to form colonies in culture and change their morphology from flat to rising/piled-up with bile canaliculi (BC), which results in maturation. In this study, we examined whether SHs could express hepatic transporters with polarity, whether the transporters could transport organic anion substrates into BC, and whether the secreted substances could be recovered from BC. Immunocytochemistry and RT-PCR were carried out. [(3)H]-labeled estrogen derivatives were used to measure the functions of the transporters in SHs isolated from normal and multidrug resistance-associated protein (Mrp) 2-deficient rats. The results showed that organic anion-transporting proteins (Oatps) 1 and 2, Na(+)-dependent taurocholate co-transporting polypeptide (Ntcp), Mrp2, and bile-salt export pump (Bsep) were well expressed in rising/piled-up cells and that their expression was correlated to that of hepatocyte nuclear factor 4alpha. Although small SHs expressed not Oatps and Mrp2 but Mrp3, rising/piled-up SHs expressed Oatp1 and 2 and Mrp2 proteins in the sinusoidal and BC membranes, respectively. On the other hand, breast cancer resistant protein (Bcrp) and Mrp3 expression decreased as SHs matured. The substrate transported via Oatps and Mrp2 was secreted into BC and it accumulated in both BC and cyst-like structures. The secreted substrate could be efficiently recovered from BC reconstructed by SHs derived from a normal rat, but not from an Mrp2-deficient rat. In conclusion, SHs can reconstitute hepatic organoids expressing functional organic anion transporters in culture. This culture system may be useful to analyze the metabolism and excretion mechanisms of drugs.

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Section: Discussionmentioning

confidence: 81%

Functional expression of organic anion transporters in hepatic organoids reconstructed by rat small hepatocytes

Oshima

Kon

Ooe

et al. 2007

J of Cellular Biochemistry

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“…A number of studies have previously investigated the in vitro hepatocyte differentiation in both mammals and freshwater fish (Ostrander et al 1995;Michalopoulos et al 1999;Mitaka et al 1999;Sidler Pfandler et al 2004;Yanhong et al 2008). Although, a number of studies on hepatocyte cultures have been carried out in freshwater species since long time, only discrete research has been performed on seabream liver, and none has been able to produce functionally active long cultured SaHePs expressing a mature phenotype (Mosconi et al 2002;Carnevali et al 2005;Bevelander et al 2006).…”

Section: Discussionmentioning

confidence: 99%

“…The expression of polarized transporters has been successfully investigated in rat SHs. These were found to be able to maintain a fully differentiated transporter expression phenotype during their transition into mature hepatocytes (Sidler Pfandler et al 2004). Here, we demonstrated that long term cultured SaHePs regain their specialized cytoarchitecture, while expressing a polarized phenotype.…”

Section: Discussionmentioning

confidence: 99%

Expression of a highly differentiated phenotype and hepatic functionality markers in gilthead seabream (Sparus aurata L.) long-cultured hepatocytes: first morphological and functional in vitro characterization

Santacroce

Zacchino

Casalino

et al. 2010

Rev Fish Biol Fisheries

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The development of primary cultures and cell lines from aquatic organisms is a valuable tool for a wide range of research activities applied to aquaculture. Despite several efforts, derivation and long-term culturing of primary hepatocytes from marine vertebrates are still rare and unsuccessful. This is the first report to fully characterize long-term cultures of primary hepatocytes from the European seabream, Sparus aurata L. (Osteichthyes, Sparidae) (SaHePs). In this new model, hepatocyte cells were long-term viable, active proliferating, and fully retained liver function up to 3 weeks. SaHePs expressed a differentiated phenotype, owing to the reacquisition of the peculiar cytoarchitecture with the complete assembly of cytoskeletal and junctional network, as shown by the production and immunolocalization of several polarity markers and cytoskeletal proteins (MDR1, ZO-2, C-CAM1, Vimentin, Cadherin, b-Tubulin, b-Catenin, b-Actin). Cytostructural analysis to identify polarized expression and bile canaliculi formation was performed by immunofluorescence and contrast phase microscopy. Long cultured SaHePs also demonstrated evidence of Albumin, a1-Antitrypsin (AAT) and a-Fetoprotein (AFP) synthesis, expression of the detoxifying metabolic enzyme cytochrome P-4501A (CYP 1A), and production of hepatocyte specific cytoskeleton proteins, such as Cytokeratin 8 (CK8) and Cytokeratin 18 (CK 18). The presence of specific markers for hepatic phenotype, detected by immunocytochemistry and Western blot analysis, is suggestive of the full maintenance of a highly differentiated phenotype and hepatic maturation. These data demonstrate that SaHePs can be long cultured without losing the hepatic functionality. This study provides a useful tool for innovative research applications in fish toxicological, pathological, and physiological studies, as one of the few hepatic, functionally active, in vitro model from marine fish.

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“…In rat small hepatocytes, which are hepatocyte progenitor cells of the adult liver, Mrp1 mRNA levels are increased approximately 2-fold during 5–9 weeks in culture (Sidler Pfandler et al, 2004). Similarly, MRP1 mRNA expression is higher in hepatic oval cells than rat hepatocytes (Zhang et al, 2007a).…”

Section: Regulation Of Hepatic Abcc1/mrp1 Transporter By Xenobiotics mentioning

confidence: 99%

Regulation of hepatic ABCC transporters by xenobiotics and in disease states

Manautou

2010

Drug Metabolism Reviews

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The subfamily of ABCC transporters consists of 13 members in mammals, including the multidrug resistance-associated proteins (MRPs), sulfonylurea receptors (SURs), and the cystic fibrosis transmembrane conductance regulator (CFTR). These proteins play roles in chemical detoxification, disposition, and normal cell physiology. ABCC transporters are expressed differentially in the liver and are regulated at the transcription and translation level. Their expression and function are also controlled by post-translational modification and membrane-trafficking events. These processes are tightly regulated. Information about alterations in the expression of hepatobiliary ABCC transporters could provide important insights into the pathogenesis of diseases and disposition of xenobiotics. In this review, we describe the regulation of hepatic ABCC transporters in humans and rodents by a variety of xenobiotics, under disease states and in genetically modified animal models deficient in transcription factors, transporters, and cell-signaling molecules.

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Small hepatocytes in culture develop polarized transporter expression and differentiation

Abstract: SummarySmall hepatocytes in culture develop polarized transporter expression and differentiation

Cited by 33 publications

References 48 publications

Functional expression of organic anion transporters in hepatic organoids reconstructed by rat small hepatocytes

Functional expression of organic anion transporters in hepatic organoids reconstructed by rat small hepatocytes

Expression of a highly differentiated phenotype and hepatic functionality markers in gilthead seabream (Sparus aurata L.) long-cultured hepatocytes: first morphological and functional in vitro characterization

Regulation of hepatic ABCC transporters by xenobiotics and in disease states

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