Arnaout W S, Moscioni A D, Barbour R L, Demetriou A A
Department of Surgery, Vanderbilt University School of Medicine, Nashville, Tennessee 37232.
J Surg Res. 1990 Apr;48(4):379-82. doi: 10.1016/0022-4804(90)90079-h.
We developed a novel bioartificial liver using microcarrier-attached hepatocytes and a bioreactor. The bioreactor consists of an intracapillary chamber made up of porous (0.2 micron) cellulose acetate hollow fibers enclosed in a polycarbonate module forming an extracapillary chamber. Cryopreserved (-80 degrees C, 3 weeks) microcarrier-attached hepatocytes (3-4 X 10(7] were inoculated into the extracapillary chamber. The bioartificial liver, containing either microcarrier hepatocytes (n = 6) or microcarriers alone (n = 5); was attached to adult Gunn rats via cannulas in the aorta and inferior vena cava. Bile samples were collected before and at hourly intervals after attachment to the bioartificial liver. The same animal model was used to demonstrate de novo conjugation of free bilirubin by the bioartificial liver (n = 3) following systemic administration of unconjugated [3H]bilirubin. Bile was analyzed for bilirubin mono- and diglucuronides by high-performance liquid chromatography and confirmed by digestion with beta-glucuronidase and cochromatography with normal rat bile. A progressive increase in the concentration of bilirubin monoglucuronide and bilirubin diglucuronide in bile was seen as early as 30 min and lasted up to 4 hr following attachment to a bioartificial liver containing microcarrier hepatocytes (3.53 +/- 0.68 to 8.07 +/- 0.85 microM; P less than 0.01). Approximately 22% of the radiolabeled bilirubin excreted in bile was seen in the form of bilirubin conjugates, no radioactivity was seen in the free bilirubin fraction. In conclusion, we developed a bioartificial liver using microcarrier hepatocytes which carry out an important differentiated liver function which results in partial correction of a specific metabolic liver defect, i.e., conjugation of bilirubin.
我们利用微载体附着的肝细胞和生物反应器开发了一种新型生物人工肝。该生物反应器由一个毛细管内腔室组成，该腔室由多孔（0.2微米）醋酸纤维素中空纤维制成，封装在聚碳酸酯模块中形成一个毛细管外腔室。将冷冻保存（-80摄氏度，3周）的微载体附着肝细胞（3 - 4×10⁷）接种到毛细管外腔室中。含有微载体肝细胞（n = 6）或仅含微载体（n = 5）的生物人工肝，通过主动脉和下腔静脉的插管与成年冈恩大鼠相连。在连接到生物人工肝之前及之后每小时采集胆汁样本。使用相同的动物模型来证明在全身给予未结合的[³H]胆红素后，生物人工肝（n = 3）对游离胆红素的从头结合。通过高效液相色谱法分析胆汁中的胆红素单葡萄糖醛酸酯和双葡萄糖醛酸酯，并通过用β - 葡萄糖醛酸酶消化以及与正常大鼠胆汁共色谱法进行确认。早在连接到含有微载体肝细胞的生物人工肝后30分钟，胆汁中胆红素单葡萄糖醛酸酯和胆红素双葡萄糖醛酸酯的浓度就开始逐渐增加，并持续长达4小时（从3.53±0.68微摩尔增加到8.07±0.85微摩尔；P小于0.01）。胆汁中排泄的放射性标记胆红素约22%以胆红素结合物的形式出现，游离胆红素部分未检测到放射性。总之，我们开发了一种使用微载体肝细胞的生物人工肝，其能够执行一项重要的分化肝功能，从而部分纠正特定的代谢性肝缺陷，即胆红素结合。
