Clinical advancement of the bioartificial liver is hampered by the lack of expandable human hepatocytes and appropriate bioreactors and carriers to encourage hepatic cells to function during extracorporeal circulation. We have recently developed an efficient approach for derivation of expandable liver progenitor-like cells from human primary hepatocytes (HepLPCs). Here, we generated immortalized and functionally enhanced HepLPCs by introducing FOXA3, a hepatocyte nuclear factor that enables potentially complete hepatic function. When cultured on macroporous carriers in an air-liquid interactive bioartificial liver (Ali-BAL) support device, the integrated cells were alternately exposed to aeration and nutrition and grew to form high-density three-dimensional constructs. This led to highly efficient mass transfer and supported liver functions such as albumin biosynthesis and ammonia detoxification via ureagenesis. In a porcine model of drug overdose–induced acute liver failure (ALF), extracorporeal Ali-BAL treatment for 3 hours prevented hepatic encephalopathy and led to markedly improved survival (83%, n = 6) compared to ALF control (17%, n = 6, P = 0.02) and device-only (no-cell) therapy (0%, n = 6, P = 0.003). The blood ammonia concentrations, as well as the biochemical and coagulation indices, were reduced in Ali-BAL–treated pigs. Ali-BAL treatment attenuated liver damage, ameliorated inflammation, and enhanced liver regeneration in the ALF porcine model and could be considered as a potential therapeutic avenue for patients with ALF.

缺乏可扩展的人类肝细胞以及合适的生物反应器和载体以促进肝细胞在体外循环中发挥功能，阻碍了生物人工肝的临床发展。我们最近开发了一种有效的方法，用于从人原代肝细胞（HepLPCs）衍生可扩展的肝祖细胞样细胞。在这里，我们通过引入FOXA3生成了永生化且功能增强的HepLPC，是一种肝细胞核因子，可以潜在地完成肝功能。当在气液交互式生物人工肝（Ali-BAL）支持设备中的大孔载体上培养时，整合细胞交替暴露于通气和营养中，并生长形成高密度的三维结构。这导致了高效的质量转移并支持了肝功能，例如白蛋白的生物合成和通过尿素生成的氨解毒。在药物过量导致的急性肝衰竭（ALF）的猪模型中，与ALF对照（17％，n）相比，体外Ali-BAL治疗3小时可预防肝性脑病并显着提高生存率（83％，n = 6）。= 6，P = 0.02）和仅设备（无细胞）疗法（0％，n = 6，P = 0.003）。经Ali-BAL处理的猪血氨浓度以及生化和凝血指标均降低。在ALF猪模型中，Ali-BAL治疗可减轻肝脏损伤，减轻炎症并增强肝脏再生，可以被视为ALF患者的潜在治疗途径。