Sandwich-cultured hepatocytes: utility forin vitroexploration of hepatobiliary drug disposition and drug-induced hepatotoxicity

Abstract

When human (cryopreserved) hepatocytes are used to establish sandwich cultures, the model appears particularly valuable to quantitatively investigate clinically relevant mechanisms related to in vivo hepatobiliary drug disposition and hepatotoxicity. Nonetheless, the SCH model would largely benefit from better insight into the fundamental cell signaling mechanisms that are critical for long-term in vitro maintenance of the hepatocytic phenotype. Studies systematically exploring improved cell culture conditions (e.g., co-cultures or extracellular matrix modifications), as well as in vitro work identifying key transcription factors involved in hepatocyte differentiation are currently emerging.

EURL ECVAM strategy for achieving 3Rs impact in the assessment of toxicokinetics and systemic toxicity

“…To our knowledge, there are no in vitro methods available at the moment for urinary excretion because of the complex renal mechanism of formation of primary urine, passive and active reuptake and the dependence of these mechanisms on differences in pressure and osmolarity between blood and primary urine. For biliary clearance, some in vitro methods have been reported (De Bruyn et al, 2013), but further development and standardisation work is necessary to identify representative methods based on which harmonised standards can be established. Although passive excretion has been suggested to suffice for 'Tier 1 PBTK modelling', it is clear that in the future, excretion based on active transporters needs to be taken into account as well.…”

Prediction of In Vivo Rat Biliary Drug Clearance from an In Vitro Hepatocyte Efflux Model

“…However, because isolated hepatocytes lose their tissue geometry, active efflux and biliary secretion of compounds are properties difficult to study in vitro (Li et al, 2010;De Bruyn et al, 2013). Robust in vitro models that also reflect biliary efflux in addition to hepatic uptake and metabolism would be valuable tools to evaluate drug disposition as well as the in vitro to in vivo extrapolation (IVIVE) of hepatic clearance.…”

“…The excretion of compounds into the canalicular compartment is used to calculate the biliary excretion (Liu et al, 1999;De Bruyn et al, 2013). This strategy has been successful in predicting the bile efflux of several drugs, but as noted by Li et al (2010) and others, it often underpredicts biliary clearance by 10-fold to 100-fold (Nakakariya et al, 2012;De Bruyn et al, 2013).…”

“…The excretion of compounds into the canalicular compartment is used to calculate the biliary excretion (Liu et al, 1999;De Bruyn et al, 2013). This strategy has been successful in predicting the bile efflux of several drugs, but as noted by Li et al (2010) and others, it often underpredicts biliary clearance by 10-fold to 100-fold (Nakakariya et al, 2012;De Bruyn et al, 2013). Partly this may be due to the down-regulation of both dx.doi.org/10.1124/dmd.113.054155. drug uptake and efflux transporters that has been demonstrated to take place within the first days after the plating of hepatocytes from both rats and humans (Liu et al, 1998;Li et al, 2009a;Kotani et al, 2011;Tchaparian et al, 2011;Kimoto et al, 2012;De Bruyn et al, 2013).…”

“…This strategy has been successful in predicting the bile efflux of several drugs, but as noted by Li et al (2010) and others, it often underpredicts biliary clearance by 10-fold to 100-fold (Nakakariya et al, 2012;De Bruyn et al, 2013). Partly this may be due to the down-regulation of both dx.doi.org/10.1124/dmd.113.054155. drug uptake and efflux transporters that has been demonstrated to take place within the first days after the plating of hepatocytes from both rats and humans (Liu et al, 1998;Li et al, 2009a;Kotani et al, 2011;Tchaparian et al, 2011;Kimoto et al, 2012;De Bruyn et al, 2013). Sinusoidal efflux transporters, such as rat multidrug resistance-associated protein 3 (rMRP3) and rat organic solute transporter a-organic solute transporter b (rOSTa-OSTb) show low expression levels in normal livers but are up-regulated in cholestasis (Rippin et al, 2001;Luttringer et al, 2002;Boyer et al, 2006).…”