Fremond B, Malandain C, Guyomard C, Chesne C, Guillouzo A, Campion J P
INSERM U 49, Hôpital Pontchaillou, Rennes.
Chirurgie. 1992;118(10):672-7.
A new type of bioartificial liver using isolated hepatocytes immobilized in alginate beads was developed and its capacity of correcting the metabolic deficiency of bilirubin conjugation in Gunn rats was assessed. Hepatocytes were isolated from Sprague-Dawley rats using the in situ collagenase perfusion technique, and they were then immobilized in Calcium alginate beads. The capacity of these immobilized hepatocytes to conjugate in vitro bilirubin was checked as compared to monolayer hepatocyte culture. The bioartificial liver consisted in a cylindrical bioreactor containing either alginate beads and hepatocytes (test group), or only alginate beads (control group). Gunn rats were connected to this bioreactor through and extracorporeal circulation system, and "bile samples were collected" every hour. Bilirubin mono and biconjugates were dosed in the bile using the high-performance liquid chromatography technique. The viability of alginate immobilized hepatocytes, determined before and after each experiment, was stable at 75%. In the test group, the total conjugate concentration rapidly increased to reach a maximum value of 204 +/- 16 microns after 3 hours, while in the control group, there were only conjugate traces (1 micron). These results show that the bioartificial liver is an efficient means to temporarily correct genetic deficiency in Gunn rats. Such a system could be of therapeutic interest in case of acute hepatic insufficiency.
一种新型的生物人工肝被研发出来，它使用固定在藻酸盐珠中的分离肝细胞，并评估了其纠正Gunn大鼠胆红素结合代谢缺陷的能力。使用原位胶原酶灌注技术从Sprague-Dawley大鼠中分离肝细胞，然后将它们固定在藻酸钙珠中。与单层肝细胞培养相比，检查了这些固定化肝细胞在体外结合胆红素的能力。生物人工肝由一个圆柱形生物反应器组成，该反应器包含藻酸盐珠和肝细胞（试验组），或仅包含藻酸盐珠（对照组）。Gunn大鼠通过体外循环系统连接到该生物反应器，每小时“收集胆汁样本”。使用高效液相色谱技术将胆红素单结合物和双结合物注入胆汁中。每次实验前后测定的藻酸盐固定化肝细胞的活力稳定在75%。在试验组中，总结合物浓度迅速增加，在3小时后达到最大值204±16微米，而在对照组中，只有痕量结合物（1微米）。这些结果表明，生物人工肝是暂时纠正Gunn大鼠遗传缺陷的有效手段。这种系统在急性肝功能不全的情况下可能具有治疗意义。
