Experimental study for reconstruction of portal vein and inferior vena cava in liver surgery

结果：2001年，携带肝细胞生长因子基因(AdHGF)的腺病毒载体在293细胞中扩增，纯化后浓缩。从犬颈静脉分离血管内皮细胞，在M199+10%胎牛血清中培养。研究AdHGF的细胞毒性和HGF的产生，以确定AdHGF的最有效剂量。结果：2003年，5只犬右肝切除后，用携带AdHGF基因的血管内皮细胞种植的PTFE移植物替代下腔静脉。5只狗中有2只活了下来，但另有3只狗在手术后一周内死亡。死因包括手术造成的无法忍受的损伤和出血。存活的两只犬于术后4周处死动物，观察肝再生情况及种子移植情况。移植物上没有凝块附着在可存活的内皮细胞上。带种子下腔静脉的肝切除犬与未植入种子的切除肝犬肝再生无明显差异。术中检测血清HGF浓度。然而，所有血清样本中的HGF水平都低于可测量的水平。提示AdHGF转导的内皮细胞在移植物上的HGF表达水平可能不足以显示其生物学活性。

Results in 2001: Adenovirus vector containing hepatocyte growth factor cDNA (AdHGF) was propagated in 293 cells and concectrated with purification. Vascular endothelial cells (ECs) were isolated from canine jugular vein and cultured in M199+10%FCS. Cytotoxicity and HGF production were investigated to determine the most effective dose of AdHGF. As a result, it was concluded that 50 PFU of AdHGF per cell were the most effective dose.Results in 2003: Replacement of IVC with PTFE graft where AdHGF-transfected ECs were seeded was performed using 5 dogs after right hepatectomy. Two out of 5 dogs were survived, but three other dogs were dead within a week after the operation. Causes of death included untolerable damages and bleeding by the operation. Survived two dogs were sacrificed 4 weeks after the operation to investigate the liver regeneration and seeds grafts. There was no clot adherent to the graft where viable ECs were attached. Liver regeneration of hepatectomized dogs with seeded IVC graft was not different from hepatectomized dogs with unseeded graft. Serum concentrations of HGF were measured in the operation. However, levels of HGF in all serum samples were less than measurable ones. It is suggested that HGF expression levels from AdHGF-transfected ECs on the graft might not be enough to show biological activity.