摘要
目的 构建生物人工肝 (BAL)并探讨其对乙型重型肝炎患者血清的代谢作用。方法 两步法分离 ,旋转振荡法培养的乳猪肝细胞球形聚集体置入中空纤维生物反应器外间隙内构建成新型BAL。应用此型BAL对乙型重型肝炎患者稀释血清进行 3h的体外灌流实验。每隔 0 .5h、灌流前及灌流后分别检测灌流血清的总胆红素、总蛋白、白蛋白和球蛋白。结果 绝大多数肝细胞在培养 2 4h后都形成了球形聚集体且保持了较高的活性。循环灌流3h后 ,与无细胞的空白对照组相比 ,治疗组血清总胆红素水平显著降低 (P <0 .0 1) ,血清总蛋白和白蛋白水平均明显增高 (P <0 .0 5 )。结论 旋转振荡法是一种简便、经济的球形聚集培养肝细胞的方法。
Objective To establish a novel bioartificial liver (BAL) consisting of spheroids of porcine hepatocytes and hollow fiber bioreactor, and study its metabolic effects on the serum of severe hepatitis B. Methods Hepatocytes were isolated from pup pigs by the two step methods, then cultured as aggregate spheroids through rotation and vibration. The spheroids of hepatocytes were transferred into the shell of a hollow fiber bioreactor to set up a novel BAL. Diluted serum samples of severe hepatitis B were circulated for 3 hours in the extracorporeal circulatory system including the BAL. Every half hour, before and after circulation, serum samples were collected to assay TBIL, TP, ALB and GLB. Results Most hepatocytes had formed spheroids with high viability when cultured for 24 hours. After 3 hours' circulation, compared with control group, the serum concentration of TBIL in treament group significantly decreased ( P < 0.01 )and the serum concentrations of TP and ALB significantly increased( P < 0.05 ). Conclusion The hepatocytes can be cultured as aggregate spheroids through a convenient and economical way known as rotation and vibration method. This novel BAL is efficient to remove bilirubin from the serum of severe hepatitis B, and supply ALB for it. Thus BAL might provide effective therapy for patients with severe hepatitis B.
出处
《西安交通大学学报(医学版)》
CAS
CSCD
北大核心
2003年第3期202-204,220,共4页
Journal of Xi’an Jiaotong University(Medical Sciences)
基金
陕西省科委科技攻关项目 (No .2 0 0 0K14 G16)