Development of bioartificial liver using transgenic pig

用胶原酶消化法分离正常猪和人衰变加速因子(HDAF)转基因猪的肝细胞。肝细胞存活率约为90%，冷冻保存后存活率为70-80%。将正常和hDAF转基因猪肝细胞暴露于人补体或人血清中，并不影响其形态和功能。另一方面，尽管转hDAF基因猪肝不发生这种情况，但与健康狒狒直接交叉循环的正常猪肝却因内皮细胞损伤和体液免疫反应而溶血而导致肝功能障碍。研制了一种用于血液透析的生物人工利华中空纤维柱，并将其用于生物人工柱。墨盒中的中空纤维是由标称截留分子量为70 kDa的三醋酸酯纤维素制成的。将分离的猪肝细胞接种于中空纤维内腔。通过负载化学物评价生物反应器接种猪肝细胞的功能。负载至少10d后，这些化学物质被迅速清除，中空纤维中的肝细胞形成排列整齐的聚集体，重新建立紧密连接和小梁状结构。

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.