Development of high performance hybrid artificial liver and its evaluation in preclinical test

1.我们建立了猪肝细胞球体（模块体积18.8cm^3，细胞重量；2g，血浆总体积；140ml）灌注培养系统，并评价了肝衰竭患者血浆中球体的功能。猪肝细胞球体在人血浆中至少在培养3天后仍保持降氨活性。为了在大型动物上验证我们的人工肝脏的性能，我们制作了混合型人工肝脏模块(多毛聚氨酯泡沫（PUF）填充床，体积；380 cm ^3)，含有30 g狗肝细胞，约占狗肝脏重量的10%。当该模块应用于缺血性肝衰竭犬（体重15 kg）时，通过人工肝治疗维持血糖浓度，抑制血氨浓度升高等。这些性能与临床上已被用作肝移植桥接的Demetriou人工肝相当或稍好。作为临床前步骤，我们使用猪肝细胞球体（体积380 cm^3*2），含64 g细胞，开发了人工肝脏模块，并应用于缺血性肝衰竭猪（体重25 kg）。因此，我们可以成功地抑制血氨浓度的升高，延长生存时间。这些结果表明，我们的混合人工肝系统是一种有前途的设备，从肝功能衰竭恢复在人类医疗保健。

1.We developed a perfusion culture system of porcine hepatocyte spheroids (module volume ; 18.8cm^3, cell weight ; 2g, total plasma volume ; 140ml), and evaluated the functions of the spheroids in plasma of hepatic failure patient. Porcine hepatocyte spheroids mainatained ammonia decreasing activity in human plasma at least 3 days of culture.2.For a validation of the performance of our artificial liver using large animals, we made hybrid artificial liver module (multi-capillary polyurethane foam (PUF) packed-bed, vulume ; 380 cm ^3) containing 30 g of dog hepatocyte which corresponded about 10% of a dog liver weight. When the module was applied to ischemic liver failure dogs (body weight ; 15 kg), blood glucose concentration was maintained, increase of blood ammonia concentration was suppressed and so on by the artificial liver treatment. These performances were comparable or slightly superior than those of Demetriou's artificial liver which was already used clinically as a bridge use for liver transplantation.3.As preclinical step, we developed an artificial liver module using porcine hepatocyte spheroids (volume ; 380 cm^3*2) containing 64 g of cells, and applied to ischemic liver failure pigs (body weight ; 25 kg). Consequently, we could succeed suppression of the increase of blood ammonia concentration and extension of survival time.These results suggest that our hybrid artificial liver system is a promising equipment for recovery from hepatic failure condition in human medical care.