Shatford R A, Nyberg S L, Meier S J, White J G, Payne W D, Hu W S, Cerra F B
Department of Surgery, University of Minnesota, Minneapolis 55455.
J Surg Res. 1992 Dec;53(6):549-57. doi: 10.1016/0022-4804(92)90253-v.
We have developed a novel hepatocyte loaded hollow fiber bioreactor as a potential bioartificial liver. Freshly harvested rat hepatocytes were entrapped in a three-dimensional gel matrix within hollow fibers in a perfused bioreactor. Gel entrapment allowed cells to be cultured at high density while maintaining tissue-specific function. Hepatocyte function was evaluated in 10 bioreactors, each containing approximately 5 x 10(7) cells. Oxygen consumption averaged 0.32 pmole/cell/hr, albumin appearance averaged 0.60 pg/cell/hr, and lidocaine clearance (a measure of the P-450 activity) averaged 0.74 pg/cell/hr. Function persisted for the 7 days of the study. Electron microscopy at 7 days showed the distinctive ultrastructure of viable, differentiated hepatocytes: bile canaliculi, intercellular junctions, peroxisomes, abundant mitochondria, and glycogen granules. Maintenance of tissue specific function and ultrastructure suggests that this bioreactor configuration has potential as a device to support patients in liver failure, as well as to study hepatocytes in vitro.
我们开发了一种新型的负载肝细胞的中空纤维生物反应器，作为一种潜在的生物人工肝。将刚收获的大鼠肝细胞包埋在灌注生物反应器中空纤维内的三维凝胶基质中。凝胶包埋使细胞能够在保持组织特异性功能的同时进行高密度培养。在10个生物反应器中评估了肝细胞功能，每个反应器含有约5×10⁷个细胞。耗氧量平均为0.32皮摩尔/细胞/小时，白蛋白生成平均为0.60皮克/细胞/小时，利多卡因清除率（一种衡量P-450活性的指标）平均为0.74皮克/细胞/小时。在研究的7天内功能持续存在。7天时的电子显微镜检查显示了存活、分化的肝细胞独特的超微结构：胆小管、细胞间连接、过氧化物酶体、丰富的线粒体和糖原颗粒。组织特异性功能和超微结构的维持表明，这种生物反应器配置有潜力作为一种支持肝衰竭患者的装置，以及在体外研究肝细胞。
