A long-term three dimensional liver co-culture system for improved prediction of clinically relevant drug-induced hepatotoxicity

Kostadinova, Radina; Boess, Franziska; Applegate, Dawn; Suter, Laura; Weiser, Thomas; Singer, Thomas P.; Naughton, Brian A.; Roth, Adrian

doi:10.1016/j.taap.2013.01.012

Cited by 237 publications

(257 citation statements)

References 92 publications

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“…Numerous advanced liver models, which improve the long-term functional performance of hepatocytes compared with monocultures, have been described (Groothuis and Meijer, 1992;LeCluyse et al, 2012); a subset of these incorporates liver nonparenchymal cells in coculture Milosevic et al, 1999;Zinchenko et al, 2006;Dash et al, 2009;Domansky et al, 2010;Kostadinova et al, 2013). For inflammatory models, cultures containing Kupffer cellseither added as a separate fraction or as part of a whole nonparenchymal fraction-are most relevant.…”

Section: Introductionmentioning

confidence: 99%

“…For inflammatory models, cultures containing Kupffer cellseither added as a separate fraction or as part of a whole nonparenchymal fraction-are most relevant. In these systems, the nonparenchymal cell fraction or Kupffer cells may be mixed with hepatocytes to form heterogeneous aggregates (Drewitz et al, 2011;Messner et al, 2013;Thoma et al, 2014), supported on a scaffold (Dash et al, 2009;Kostadinova et al, 2013), or separated through micropatterning (Zinchenko et al, 2006). Transwell devices have also been evaluated .…”

Section: Introductionmentioning

confidence: 99%

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Metabolite Profiling and Pharmacokinetic Evaluation of Hydrocortisone in a Perfused Three-Dimensional Human Liver Bioreactor

et al. 2015

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Endotoxin lipopolysaccharide (LPS) is known to cause liver injury primarily involving inflammatory cells such as Kupffer cells, but few in vitro culture models are applicable for investigation of inflammatory effects on drug metabolism. We have developed a threedimensional human microphysiological hepatocyte-Kupffer cell coculture system and evaluated the anti-inflammatory effect of glucocorticoids on liver cultures. LPS was introduced to the cultures to elicit an inflammatory response and was assessed by the release of proinflammatory cytokines, interleukin 6 and tumor necrosis factor a. A sensitive and specific reversed-phase-ultra high-performance liquid chromatography-quadrupole time of flightmass spectrometry method was used to evaluate hydrocortisone disappearance and metabolism at near physiologic levels. For this, the systems were dosed with 100 nM hydrocortisone and circulated for 2 days; hydrocortisone was depleted to approximately 30 nM, with first-order kinetics. Phase I metabolites, including tetrahydrocortisone and dihydrocortisol, accounted for 8-10% of the loss, and 45-52% consisted of phase II metabolites, including glucuronides of tetrahydrocortisol and tetrahydrocortisone. Pharmacokinetic parameters, i.e., half-life, rate of elimination, clearance, and area under the curve, were 23.03 hours, 0.03 hour 21 , 6.6 3 10 25 l×hour 21, and 1.03 (mg/l)*h, respectively. The ability of the bioreactor to predict the in vivo clearance of hydrocortisone was characterized, and the obtained intrinsic clearance values correlated with human data. This system offers a physiologically relevant tool for investigating hepatic function in an inflamed liver.

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Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Metabolite Profiling and Pharmacokinetic Evaluation of Hydrocortisone in a Perfused Three-Dimensional Human Liver Bioreactor

et al. 2015

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“…2 Thus, the advantages of such cultures are (1) drugs that have been identified as false-negatives would be identified as effective and (2) late drug failure and possibly even drug removal from the market could be avoided, and thus a lot of money could be saved. 3 Three-dimensional cell culture opens new dimensions in cell-based assays. 4 For screening drugs, 3D cell cultures must be automated to obtain appropriate throughput and must be well reproducible and standardizable, 5 the latter being also important to obtain approval by the authorities.…”

Section: Introductionmentioning

confidence: 99%

Automation of 3D Cell Culture Using Chemically Defined Hydrogels

Rimann

Angres²,

Patocchi-Tenzer

et al. 2014

SLAS Technology

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“…[29][30][31][32][33][34][35][36][37][38] Several strategies have previously been proposed to culture LSECs while retaining their phenotype. [37][38][39][40][41][42][43] LSECs, cultured in media supplemented with lipids and oleic acid, in particular, was shown to maintain their metabolic and endocytotic activity for up to 5 days by influencing Akt/PKB and ERK signaling pathways. 39 Another study with culturing LSECs and hepatocytes in a three-dimensional (3D) microreactor shows retention of LSEC phenotype expression for 13 days, primarily through modulation of vascular endothelial growth factor in the culture media.…”

mentioning

confidence: 99%

Long-Term Coculture Strategies for Primary Hepatocytes and Liver Sinusoidal Endothelial Cells

Bale

Golberg

Jindal

et al. 2015

Tissue Engineering Part C: Methods

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Hepatocytes and their in vitro models are essential tools for preclinical screening studies for drugs that affect the liver. Most of the current models primarily focus on hepatocytes alone and lack the contribution of nonparenchymal cells (NPCs), which are significant through both molecular and the response of the NPCs themselves. Models that incorporate NPCs alongside hepatocytes hold the power to enable more realistic recapitulation and elucidation of cell interactions and cumulative drug response. Hepatocytes and liver sinusoidal endothelial cells (LSECs) account for *80% of the liver mass where the LSECs line the walls of blood vessels, and act as a barrier between hepatocytes and blood. Culturing LSECs with hepatocytes to generate multicellular physiologically relevant in vitro liver models has been a major hurdle since LSECs lose their phenotype rapidly after isolation. To this end, we describe the application of collagen gel (1) in a sandwich and (2) as an intervening extracellular matrix layer to coculture hepatocytes with LSECs for extended periods. These coculture configurations provide environments wherein hepatocyte and LSECs, through cell-cell contacts and/or secretion factors, lead to enhanced function and stability of the cocultures. Our results show that in these configurations, hepatocytes and LSECs maintained their phenotypes when cultured together as a mixture, and showed stable secretion and metabolic activity for up to 4 weeks. Immunostaining for sinusoidal endothelial 1 (SE-1) antibody demonstrated retention of LSEC phenotype during the culture period. In addition, LSECs cultured alone maintained high viability and SE-1 expression when cultured within a collagen sandwich configuration up to 4 weeks. Albumin production of the cocultures was 10-15 times higher when LSECs were cultured as a bottom layer (with an intervening collagen layer) and as a mixture in a sandwich configuration, and native CYP 1A1/2 activity was at least 20 times higher than monoculture controls. Together, these data suggest that collagen gel-based hepatocyte-LSEC cocultures are highly suitable models for stabilization and long-term culture of both cell types. In summary, these results indicate that collagen gel-based hepatocyte-LSEC coculture models are promising for in vitro toxicity testing, and liver model development studies.

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A long-term three dimensional liver co-culture system for improved prediction of clinically relevant drug-induced hepatotoxicity

Cited by 237 publications

References 92 publications

Metabolite Profiling and Pharmacokinetic Evaluation of Hydrocortisone in a Perfused Three-Dimensional Human Liver Bioreactor

Metabolite Profiling and Pharmacokinetic Evaluation of Hydrocortisone in a Perfused Three-Dimensional Human Liver Bioreactor

Automation of 3D Cell Culture Using Chemically Defined Hydrogels

Long-Term Coculture Strategies for Primary Hepatocytes and Liver Sinusoidal Endothelial Cells

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