经肝静脉与门静脉离体灌注分离猪肝细胞的比较研究  

A comparitive study of perfusion via portal vein and perfusion via hepatic vein in porcine hepatocyte isolation

作　　者：李涛[1] 彭志海[1] 孙星[1] 唐华美[2] 裘国强[1] 

机构地区：[1]上海市第一人民医院普外科,200080 [2]上海市第一人民医院病理科,200080

出　　处：《肝脏》2005年第1期22-23,共2页Chinese Hepatology

摘　　要：目的　比较经门静脉顺行灌注法与肝静脉逆行灌注法离体分离屠宰场猪肝细胞并观察原代培养的肝细胞形态学变化。方法　经门静脉顺行和肝静脉逆行离体胶原酶灌注法分离猪肝细胞 ,并在含 10 %小牛血清的WilliamsE培养基中培养。结果　顺行灌注与逆行灌注法分离猪肝细胞的平均产量分别为 (8.8± 0 .5 )× 10 9 肝与 (1.5± 0 .1)× 10 1 0 肝 (P <0 .0 5 ) ,平均肝细胞活性为 (88.7± 1.5 ) %与 (90 .3± 1.5 ) % (P >0 .0 5 )。分离后的肝细胞增生活跃 ,生长良好 ,未见污染。结论　采用肝静脉逆行灌注法可以分离获得高产率、高活性肝细胞 。Objective To compare the isolation methods for collection of porcine hepatocytes. Methods Two-step collagenase digestion method via portal vein or hepatic vein was used to isolate porcine hepatocytes and isolated hepatocytes were cultured in the Williams E media supplemented with 10% calf serum.Results The hepatocytes yield was significantly higher (P<0.05) in the hepatic venous perfusion group, averaging(8.8±0.5)×10 9/liver in the portal venous perfusion group and (1.5±0.1)×10 (10)/liver in the hepatic venous perfusion group, respectively. Comparison of the viability showed no significant differences (P>0.05) between two groups, (88.7±1.5)% and (90.3±1.5)% in the portal venous perfusion group and the hepatic venous perfusion group, respectively. Cultured porcine hepatocytes showed high proliferative ability and grew well.Conclusions The retrograde perfusion via hepatic vein allows isolation of large numbers of viable pig hepatocytes, this can provide sufficient hepatocytes for bioartificial liver.

关 键 词：猪肝细胞 分离 离体灌注 经肝静脉 WILLIAMS 胶原酶灌注法 经门静脉 形态学变化 肝细胞活性 生物人工肝 原代培养 小牛血清 顺行灌注 细胞来源 逆行 屠宰场 培养基 细胞增 高活性 平均 

分 类 号：R329.2[医药卫生—人体解剖和组织胚胎学]