In vitro transport studies across cells grown on culture inserts are widely used for evaluating pharmacokinetic characteristics such as intestinal membrane permeability. However, measurements of the apparent permeability coefficient of highly lipophilic compounds are often limited by transport across the membrane filters, not by transport across the cultured cells. To overcome this concern, we have investigated the utility of a high-porosity membrane honeycomb film (HCF) for transcellular transport studies. Using the HCF inserts, the apparent permeability coefficient (P app ) of the drugs tested in LLC-PK1 and Caco-2 cells tended to increase with an increase in lipophilicity, reaching a maximum P app value at Log D higher than 2. In contrast, using the commercially available Track-Etched membrane (TEM) inserts, a maximum value was observed at Log D higher than 1. The basolateral to apical transport permeability P app(BL→AP) of rhodamine 123 across LLC-PK1 cells that express P-glycoprotein (P-gp) cultured on HCF inserts and TEM inserts was 2.33 and 2.39 times higher than the reverse directional P app(AP→BL) permeability, respectively. The efflux ratio (P app(B-A) / P app(A-B) ) of rhodamine 123 in LLC-PK1 expressing P-gp cells using HCF inserts was comparable to that obtained using TEM inserts, whereas the transported amount in both directions was significantly higher when using the HCF inserts. Accordingly, due to the higher permeability and high porosity of HCF membranes, it is expected that transcellular transport of high lipophilic as well as hydrophilic compounds and substrate recognition of transporters can be evaluated more accurately by using HCF inserts.

No abstract

No abstract

No abstract

Biliary excretion via drug-metabolizing enzymes and transporters is often critical as the hepatobiliary disposition is directly linked to drug efficacy and safety. Sandwich-cultured hepatocyte (SCH) is an in vitro model widely used to assess the biliary excretion potential of drugs. However, SCH does not allow time-dependent and quantitative evaluation because drugs accumulate in the closed bile canaliculi formed between hepatocytes and their amounts cannot be measured directly. This structural problem of SCH can be overcome by inducing a formation of an open canaliculus lumen face to a culture support. The aim of this study is to establish a novel hepatocyte culture model with open-form bile canaliculi that enables to evaluate biliary excretion of drugs by a permeation assay but not by accumulation assay.