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[Development of novel culture system for regulation of hepatocyte function].

Cultured hepatocytes are expected to be used for drug screening and bioartificial liver. Since hepatocytes lose their functions very rapidly in vitro, many attempts have been made to maintain their viability and functions. First, we want to introduce the surface modification of culture substrate using a starburst dendrimer. Addition of fructose to the terminal of the dendrimer was shown to be effective in maintaining hepatocyte function. As the second topic, we will show results of the use of a three-dimensional carrier for hepatocyte cultivation. Hepatocytes and bone marrow stromal cells were cocultured in silane beads, and packed into a radial flow-type bioreactor. The perfusion culture showed the effectiveness of bone marrow stromal cells for the maintenance of hepatocyte function. The next topic will be the trial of adenoviral gene transfer into hepatocytes. Thioredoxin gene was chosen because the products play important roles in redox control and antiapoptosis. The introduction of the gene could inhibit apoptosis and maintain the hepatocyte viability. Finally, we want to introduce the results on differentiation of stem cells into hepatocytes, because it is very difficult to obtain sufficient number of human hepatocytes. Human mesenchymal stem cells were cultured in the presence of several protein factors and the hepatocyte-specific marker was expressed after 2 weeks of induction culture. The use of human stem cells could be an important strategy for the support of a drug development system.