Establishment of simulation model for human drug metabolism by bioartificial liver

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We have continued to study by using radial flow bio artificial liver. The expression of CYP3A4 was extremely increased after culturing liver cells in this system. We used FLC cells which originally expressed CYP3A4 and keep high function. After we cultured other liver cells that express CYP3A4 in RFB, CYP3A4 mRNA expressed much more than monolayer culture as well as FLC cells. Furthermore, we cultured GS3A4-HepG2 which introdued gene of glutamin synthetase and CYP3A4 in RFB. As a result, we determined that annmonium was metabolized effectively by those cells under RFB culture.Next, we investigated whether other several liver cell lines are suitable for RFB culture or not. We got the results that the cells which those doubling time were shorter than 24hr could culture on RFB. We speculated that the cells which doubling time was longer than 24hr might be thrown away before attaching on culture matrix. We presented these results at the Japanese congress of regeneration on 2005.On the othe … More r hand, we investigated the differences of expression of CYP1A1, 2D6, 2A6, 2C9, and 2C19 except for CYP3A4 after culturing on RFB or monolayer. As a result, we only got the differences of expression of 2D6 after culturing between them. We could not get the differences of 1A1, which means that enzyme associated with tumorgenesis did not induce.But, expression of arginosuccinate lyase, carbamoyl-p-synthetase, argininosuccinate synthetase that are uremic enzyme on those cell lines increased after RFB culture, and their production of alphafetoprotein (AFP) was inhibited.Furthermore, we also tried to culture kidney cells on RFB because we want to apply RFB for treatment of hepatocellular syndrome.Among them, proximal tubular cells (PTC) were capable of culture on RFB and their function also was increased. For example, expression of CYP27 was increased and expression extracellular matrix was increased on high glucose/hypoxic culture of PTC by using RFB. These results indicated that both high glucose and hypoxia might be necessary to induce acute renal failure. We presented these results at the congress of regeneration on 2005 and congress of Tokyo medical university on 2006, and Dr.Jojima got the grant from Tokyo medical university. Finally, we reported these results on Journal of Tokyo medical university on 2006. Less