A tissue engineering approach for liver tissue organization in vitro

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We previously described the mass preparation ofhepatocyte spheroids in a cell suspension. Addition ofa synthetic polymer, Eudragit S 100 (a co-polymer of methacrylic acid and methylmethacrylate), to the culture medium as an artificial matrix has been found to promote cell aggregation and induce spheroid formation. The present study examines co-cultures of hepatocytgs and liver non-parenchymal cells to further enhance the stable expression of liver functions. Hetero-spheroids consisting of "hepatocyteT.and non-parenchymal cells were also formed by adding the polymer to the culture medium. The expression levels of liver functions depended on the mixing ratio of the cells The values ofalbumin secretion, anmlonia removal and urea synthesis were the highest when equal number ofhepatocytes and non-parenchymal cells were mixed. The hetero-spherpids maintained a higher level ofalbumin secretion for over 4 weeks^compared with co-cultured cells without the polymer and spheroids consisting of hepatocytes solely.^Other liver functions such as anunonia removal and cytochrome P-450 activity in'hetero-spheroid cultures were also maintained at a high level throughout the 4-week culture period.