Limits...
Recent advances in 2D and 3D in vitro systems using primary hepatocytes, alternative hepatocyte sources and non-parenchymal liver cells and their use in investigating mechanisms of hepatotoxicity, cell signaling and ADME.

Godoy P, Hewitt NJ, Albrecht U, Andersen ME, Ansari N, Bhattacharya S, Bode JG, Bolleyn J, Borner C, Böttger J, Braeuning A, Budinsky RA, Burkhardt B, Cameron NR, Camussi G, Cho CS, Choi YJ, Craig Rowlands J, Dahmen U, Damm G, Dirsch O, Donato MT, Dong J, Dooley S, Drasdo D, Eakins R, Ferreira KS, Fonsato V, Fraczek J, Gebhardt R, Gibson A, Glanemann M, Goldring CE, Gómez-Lechón MJ, Groothuis GM, Gustavsson L, Guyot C, Hallifax D, Hammad S, Hayward A, Häussinger D, Hellerbrand C, Hewitt P, Hoehme S, Holzhütter HG, Houston JB, Hrach J, Ito K, Jaeschke H, Keitel V, Kelm JM, Kevin Park B, Kordes C, Kullak-Ublick GA, LeCluyse EL, Lu P, Luebke-Wheeler J, Lutz A, Maltman DJ, Matz-Soja M, McMullen P, Merfort I, Messner S, Meyer C, Mwinyi J, Naisbitt DJ, Nussler AK, Olinga P, Pampaloni F, Pi J, Pluta L, Przyborski SA, Ramachandran A, Rogiers V, Rowe C, Schelcher C, Schmich K, Schwarz M, Singh B, Stelzer EH, Stieger B, Stöber R, Sugiyama Y, Tetta C, Thasler WE, Vanhaecke T, Vinken M, Weiss TS, Widera A, Woods CG, Xu JJ, Yarborough KM, Hengstler JG - Arch. Toxicol. (2013)

Bottom Line: When isolating liver cells, some pathways are activated, e.g., the RAS/MEK/ERK pathway, whereas others are silenced (e.g. HNF-4α), resulting in up- and downregulation of hundreds of genes.One key message is that despite our enthusiasm for in vitro systems, we must never lose sight of the in vivo situation.Although hepatocytes have been isolated for decades, the hunt for relevant alternative systems has only just begun.

View Article: PubMed Central - PubMed

Affiliation: Leibniz Research Centre for Working Environment and Human Factors (IFADO), 44139, Dortmund, Germany.

ABSTRACT
This review encompasses the most important advances in liver functions and hepatotoxicity and analyzes which mechanisms can be studied in vitro. In a complex architecture of nested, zonated lobules, the liver consists of approximately 80 % hepatocytes and 20 % non-parenchymal cells, the latter being involved in a secondary phase that may dramatically aggravate the initial damage. Hepatotoxicity, as well as hepatic metabolism, is controlled by a set of nuclear receptors (including PXR, CAR, HNF-4α, FXR, LXR, SHP, VDR and PPAR) and signaling pathways. When isolating liver cells, some pathways are activated, e.g., the RAS/MEK/ERK pathway, whereas others are silenced (e.g. HNF-4α), resulting in up- and downregulation of hundreds of genes. An understanding of these changes is crucial for a correct interpretation of in vitro data. The possibilities and limitations of the most useful liver in vitro systems are summarized, including three-dimensional culture techniques, co-cultures with non-parenchymal cells, hepatospheres, precision cut liver slices and the isolated perfused liver. Also discussed is how closely hepatoma, stem cell and iPS cell-derived hepatocyte-like-cells resemble real hepatocytes. Finally, a summary is given of the state of the art of liver in vitro and mathematical modeling systems that are currently used in the pharmaceutical industry with an emphasis on drug metabolism, prediction of clearance, drug interaction, transporter studies and hepatotoxicity. One key message is that despite our enthusiasm for in vitro systems, we must never lose sight of the in vivo situation. Although hepatocytes have been isolated for decades, the hunt for relevant alternative systems has only just begun.

Show MeSH

Related in: MedlinePlus

Distribution of extracellular matrix (ECM) in the liver acinus. A basement membrane is localized in the periportal and perivenous regions. Fibronectin is the main ECM component of the liver parenchyma, and it is localized in the space of Dissé. Adapted from Rodés (2007)
© Copyright Policy - OpenAccess
Related In: Results  -  Collection


getmorefigures.php?uid=PMC3753504&req=5

Fig3: Distribution of extracellular matrix (ECM) in the liver acinus. A basement membrane is localized in the periportal and perivenous regions. Fibronectin is the main ECM component of the liver parenchyma, and it is localized in the space of Dissé. Adapted from Rodés (2007)

Mentions: Compared to other organs, the liver is not particularly rich in ECM. Nevertheless, the ECM plays an important role in maintaining the differentiated phenotype of hepatocytes and NPCs (Martinez-Hernandez and Amenta 1993; Schuppan et al. 2001). Significant ECM alterations are observed in liver cirrhosis and fibrosis (Schuppan et al. 2001; Wells 2008a). The phenotypic changes induced by increasing the ECM stiffness are summarized in Table 1. As expected, isolated hepatocytes de-differentiate when cultured on hard 2D substrates that increase the ECM stiffness to favor a proliferative rather than differentiated cellular phenotype (Wells 2008a, b). The ECM composition roughly follows a gradient in the region comprised between the periportal and the perivenous areas (Table S2; see 10.1007/s00204-013-1078-5). Basement membrane proteins (consisting of laminin, collagen type IV and perlecan) are mostly concentrated around the portal blood vessels and the larger venes. Here, the ECM composition is similar to that of other epithelial organs. By contrast, the basement membrane is absent in the parenchyma. The ECM in the parenchyma is located in the space of Dissé between the hepatocyte plates and the sinusoids (Fig. 3). Fibronectin and collagen I dominate in the parenchyma, with smaller amounts of collagen type III. The effect of the matrix components is striking in hepatic progenitor cells. Collagen I favors the differentiation of hepatic stem cells, while laminin maintains stemness (McClelland et al. 2008).Table 1


Recent advances in 2D and 3D in vitro systems using primary hepatocytes, alternative hepatocyte sources and non-parenchymal liver cells and their use in investigating mechanisms of hepatotoxicity, cell signaling and ADME.

Godoy P, Hewitt NJ, Albrecht U, Andersen ME, Ansari N, Bhattacharya S, Bode JG, Bolleyn J, Borner C, Böttger J, Braeuning A, Budinsky RA, Burkhardt B, Cameron NR, Camussi G, Cho CS, Choi YJ, Craig Rowlands J, Dahmen U, Damm G, Dirsch O, Donato MT, Dong J, Dooley S, Drasdo D, Eakins R, Ferreira KS, Fonsato V, Fraczek J, Gebhardt R, Gibson A, Glanemann M, Goldring CE, Gómez-Lechón MJ, Groothuis GM, Gustavsson L, Guyot C, Hallifax D, Hammad S, Hayward A, Häussinger D, Hellerbrand C, Hewitt P, Hoehme S, Holzhütter HG, Houston JB, Hrach J, Ito K, Jaeschke H, Keitel V, Kelm JM, Kevin Park B, Kordes C, Kullak-Ublick GA, LeCluyse EL, Lu P, Luebke-Wheeler J, Lutz A, Maltman DJ, Matz-Soja M, McMullen P, Merfort I, Messner S, Meyer C, Mwinyi J, Naisbitt DJ, Nussler AK, Olinga P, Pampaloni F, Pi J, Pluta L, Przyborski SA, Ramachandran A, Rogiers V, Rowe C, Schelcher C, Schmich K, Schwarz M, Singh B, Stelzer EH, Stieger B, Stöber R, Sugiyama Y, Tetta C, Thasler WE, Vanhaecke T, Vinken M, Weiss TS, Widera A, Woods CG, Xu JJ, Yarborough KM, Hengstler JG - Arch. Toxicol. (2013)

Distribution of extracellular matrix (ECM) in the liver acinus. A basement membrane is localized in the periportal and perivenous regions. Fibronectin is the main ECM component of the liver parenchyma, and it is localized in the space of Dissé. Adapted from Rodés (2007)
© Copyright Policy - OpenAccess
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3753504&req=5

Fig3: Distribution of extracellular matrix (ECM) in the liver acinus. A basement membrane is localized in the periportal and perivenous regions. Fibronectin is the main ECM component of the liver parenchyma, and it is localized in the space of Dissé. Adapted from Rodés (2007)
Mentions: Compared to other organs, the liver is not particularly rich in ECM. Nevertheless, the ECM plays an important role in maintaining the differentiated phenotype of hepatocytes and NPCs (Martinez-Hernandez and Amenta 1993; Schuppan et al. 2001). Significant ECM alterations are observed in liver cirrhosis and fibrosis (Schuppan et al. 2001; Wells 2008a). The phenotypic changes induced by increasing the ECM stiffness are summarized in Table 1. As expected, isolated hepatocytes de-differentiate when cultured on hard 2D substrates that increase the ECM stiffness to favor a proliferative rather than differentiated cellular phenotype (Wells 2008a, b). The ECM composition roughly follows a gradient in the region comprised between the periportal and the perivenous areas (Table S2; see 10.1007/s00204-013-1078-5). Basement membrane proteins (consisting of laminin, collagen type IV and perlecan) are mostly concentrated around the portal blood vessels and the larger venes. Here, the ECM composition is similar to that of other epithelial organs. By contrast, the basement membrane is absent in the parenchyma. The ECM in the parenchyma is located in the space of Dissé between the hepatocyte plates and the sinusoids (Fig. 3). Fibronectin and collagen I dominate in the parenchyma, with smaller amounts of collagen type III. The effect of the matrix components is striking in hepatic progenitor cells. Collagen I favors the differentiation of hepatic stem cells, while laminin maintains stemness (McClelland et al. 2008).Table 1

Bottom Line: When isolating liver cells, some pathways are activated, e.g., the RAS/MEK/ERK pathway, whereas others are silenced (e.g. HNF-4α), resulting in up- and downregulation of hundreds of genes.One key message is that despite our enthusiasm for in vitro systems, we must never lose sight of the in vivo situation.Although hepatocytes have been isolated for decades, the hunt for relevant alternative systems has only just begun.

View Article: PubMed Central - PubMed

Affiliation: Leibniz Research Centre for Working Environment and Human Factors (IFADO), 44139, Dortmund, Germany.

ABSTRACT
This review encompasses the most important advances in liver functions and hepatotoxicity and analyzes which mechanisms can be studied in vitro. In a complex architecture of nested, zonated lobules, the liver consists of approximately 80 % hepatocytes and 20 % non-parenchymal cells, the latter being involved in a secondary phase that may dramatically aggravate the initial damage. Hepatotoxicity, as well as hepatic metabolism, is controlled by a set of nuclear receptors (including PXR, CAR, HNF-4α, FXR, LXR, SHP, VDR and PPAR) and signaling pathways. When isolating liver cells, some pathways are activated, e.g., the RAS/MEK/ERK pathway, whereas others are silenced (e.g. HNF-4α), resulting in up- and downregulation of hundreds of genes. An understanding of these changes is crucial for a correct interpretation of in vitro data. The possibilities and limitations of the most useful liver in vitro systems are summarized, including three-dimensional culture techniques, co-cultures with non-parenchymal cells, hepatospheres, precision cut liver slices and the isolated perfused liver. Also discussed is how closely hepatoma, stem cell and iPS cell-derived hepatocyte-like-cells resemble real hepatocytes. Finally, a summary is given of the state of the art of liver in vitro and mathematical modeling systems that are currently used in the pharmaceutical industry with an emphasis on drug metabolism, prediction of clearance, drug interaction, transporter studies and hepatotoxicity. One key message is that despite our enthusiasm for in vitro systems, we must never lose sight of the in vivo situation. Although hepatocytes have been isolated for decades, the hunt for relevant alternative systems has only just begun.

Show MeSH
Related in: MedlinePlus