Development of bioartificial liver using transgenic pig

1.人工肝用转基因猪肝的形态学和功能评价用胶原酶消化法从正常猪和转人衰变加速因子（human decay accelerating factor，hEGF）基因猪中分离出猪肝细胞。肝细胞的存活率约为90%，冻存后为70-80%。正常和hDAF转基因猪肝细胞暴露于人补体或人血清对其形态和功能没有影响。另一方面，正常猪肝直接与健康狒狒交叉循环后，由于内皮细胞损伤和体液免疫反应引起的溶血而导致肝功能障碍，而转hDAF猪肝则没有.生物人工肝生物反应器的研制采用临床上用于血液透析的中空纤维滤芯作为生物人工滤芯。药筒中的中空纤维由标称截止分子量为70 kDa的三乙酸纤维素制成。将分离的猪肝细胞接种到中空纤维的内部空间中。13通过加载化学物质来评价生物反应器接种猪肝细胞的功能，加载至少10天后，这些化学物质迅速消除，中空纤维中的肝细胞形成了组织良好的聚集体，重新建立了紧密连接和小梁样结构。

1. Morpholugical and functional evaluation of transgenic pig liver for artificial liverPorcine hepatocytes were isolated from normal pigs and human decay accelerating factor (hDAF)-transgenic pigs with collagenase digestion method. The viability of hepatocytes was approximately 90%, and 70-80% after cryopreservation. Exposure of normal and hDAF-transgenic porcine hepatocytes to human complements or human serum did not affect on their morphology and function. On the other hand, normal pig livers cross-circulated directly with healthy baboons were induced liver dysfunction due to endothelial cell injury and -hemolysis by humoral immunoreactions, although hDAF-transgenic pig livers were not.2. Development of bioreactor for bioartificial liverA hollow fiber cartridge that has been clinically used for hemodialysis was used for a bioartificial cartridge. The hollow fiber in the cartridge was made of cellulose triacetate with nominal cut-off molecular weight of 70 kDa. The isolated porcine hepatocytes were inoculated into the inner space of hollow fiber. The function of bioreactor inoculated porcine hepatocytes was evaluated by loading chemicalsThese chemicals were rapidly eliminated after loading for at least 10 days, and the hepatocyte in the hollow fiber formed well-organized aggregates with re-established tight junction and trabecular like structure.