Development of bioartificial liver: bilirubin conjugation in Gunn rats.
Development of bioartificial liver: bilirubin conjugation in Gunn rats.
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生物人工肝的发育：Gunn 大鼠中的胆红素结合。
DOI：
10.1016/0022-4804(90)90079-h
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发表时间：
1990
期刊：
影响因子：
--
通讯作者：
Demetriou,AA
中科院分区：
文献类型：
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作者：
Arnaout,WS;Moscioni,AD;Barbour,RL;Demetriou,AA
我们利用微载体附着的肝细胞和生物反应器研制了一种新型的生物人工肝。生物反应器由一个毛细管内腔组成，该毛细管内腔由多孔（0.2 μm）醋酸纤维素中空纤维组成，封闭在聚碳酸酯模块中，形成一个毛细管外腔。将冻存（−80°C，3周）的微载体附着肝细胞（3-4 × 107）接种到毛细管外室中。将含有微载体肝细胞（n= 6）或仅含有微载体（n= 5）的生物人工肝通过主动脉和下腔静脉中的插管附着于成年古恩大鼠。胆汁样品收集之前和之后的每小时的时间间隔连接到生物人工肝。使用相同的动物模型来证明在全身给予未结合的[3 H]胆红素后，生物人工肝（n= 3）对游离胆红素的新结合。通过高效液相色谱法分析胆汁中的胆红素单葡萄糖醛酸和二葡萄糖醛酸，并通过β-葡萄糖醛酸酶消化和与正常大鼠胆汁的共色谱法进行确认。胆汁中胆红素单葡萄糖醛酸苷和胆红素二葡萄糖醛酸苷的浓度在附着到含有微载体肝细胞的生物人工肝后30 min就观察到进行性增加，并持续至4 h（3.53 ± 0.68至8.07 ± 0.85 μM;P< 0.01）。在胆汁中排泄的放射性标记胆红素中，约22%以胆红素结合物的形式存在，在游离胆红素部分中未观察到放射性。总之，我们开发了一种使用微载体肝细胞的生物人工肝，该微载体肝细胞进行重要的分化肝功能，其导致特定代谢性肝缺陷的部分校正，即，胆红素结合。
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 μm) cellulose acetate hollow fibers enclosed in a polycarbonate module forming an extracapillary chamber. Cryopreserved (−80°C, 3 weeks) microcarrier-attached hepatocytes (3–4 × 107) 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 demonstratede novoconjugation 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 β-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 μM;P< 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.