A tissue engineering approach for liver tissue organization in vitro

我们以前描述过在细胞悬浮液中大量制备肝细胞球体。在培养基中加入一种合成聚合物Eudragit s100（一种甲基丙烯酸和甲基丙烯酸甲酯的共聚物）作为人工基质，可以促进细胞聚集并诱导球体形成。本研究探讨肝细胞和肝非实质细胞的共培养，以进一步增强肝功能的稳定表达。由“肝细胞tet”组成的异质球体。在培养基中加入该聚合物也可形成非实质细胞。肝脏功能的表达水平与细胞的混合比例有关，当等量的肝细胞和非实质细胞混合时，白蛋白分泌、排空和尿素合成的值最高。与不含聚合物的共培养细胞和仅由肝细胞组成的球体相比，异质球体在4周内保持较高水平的白蛋白分泌。^其他肝脏功能，如在异球培养物中去除anunonia和细胞色素P-450活性在整个4周的培养期间也保持在高水平。

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.