Wang L, Sun J, Li L, Mears D, Horvat M, Sheil A G
Royal Prince Alfred Hospital and Department of Surgery, University of Sydney, NSW, Australia.
Cell Transplant. 1998 Sep-Oct;7(5):459-68. doi: 10.1177/096368979800700505.
Cells from primary porcine hepatocytes (PPH) and the immortalized human hepatoma cell line C3A are both used in bioartificial liver support systems (BALSS). In this work the viability and metabolic capacity of PPH and C3A cells cultured in different media were compared. Also, because the cells come into direct or indirect contact with human blood components in BALSS, the effects of human complement on survival and functions of the cells was evaluated. For short-term culture, maintenance of PPH viability was essential for retention of P450IA1 activity (r = 0.882, p < 0.01) and effective ammonia clearance (r = -0.791, p < 0.01). When cell viability was below 60% P450IA1 activity could not be recorded and nitrogen elimination activity significantly diminished. In contrast to PPH, ammonia levels were markedly increased for C3A cells in all culture media tested (p < 0.01). Ammonia increase correlated with C3A viability (r = 0.896, p < 0.05). PPH metabolic function was superior to that of the C3A cell line when evaluated by P450IA1 activity, ammonia removal, and amino acid metabolism. When PPH were incubated in human plasma (HP) or human serum (HS) there was rapid and irreversible deterioration of viability occurring within 9 h. This toxic effect could be prevented by the inactivation of complement. When sodium citrate dissolved in dextrose was added to medium, there was considerable damage to both PPH and the C3A cell line. However, there was no demonstrable toxic effect when hepatic cells of either type were exposed to heparin. We conclude that PPH cultivated in complement-inactivated HP or HS are to be preferred to C3A for clinical application of BALSS, and that heparin should be preferred for anticoagulation in BALSS.
原代猪肝细胞（PPH）和永生化人肝癌细胞系C3A的细胞都用于生物人工肝支持系统（BALSS）。在这项工作中，比较了在不同培养基中培养的PPH和C3A细胞的活力和代谢能力。此外，由于细胞在BALSS中与人体血液成分直接或间接接触，因此评估了人补体对细胞存活和功能的影响。对于短期培养，维持PPH活力对于保留P450IA1活性（r = 0.882，p < 0.01）和有效氨清除（r = -0.791，p < 0.01）至关重要。当细胞活力低于60%时，无法记录P450IA1活性，氮清除活性显著降低。与PPH相反，在所有测试的培养基中，C3A细胞的氨水平均显著升高（p < 0.01）。氨的增加与C3A活力相关（r = 0.896，p < 0.05）。通过P450IA1活性、氨去除和氨基酸代谢评估时，PPH的代谢功能优于C3A细胞系。当PPH在人血浆（HP）或人血清（HS）中孵育时，9小时内活力迅速且不可逆地下降。这种毒性作用可通过补体失活来预防。当将溶解在葡萄糖中的柠檬酸钠添加到培养基中时，PPH和C3A细胞系均受到相当大的损害。然而，当任何一种类型的肝细胞暴露于肝素时，均未显示出明显的毒性作用。我们得出结论，在BALSS的临床应用中，在补体失活的HP或HS中培养的PPH比C3A更适合，并且在BALSS中肝素应优先用于抗凝。
