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

结果2001年：在293细胞中获得含肝细胞生长因子cDNA （AdHGF）的腺病毒载体，并进行浓缩纯化。从犬颈静脉中分离血管内皮细胞，在M199+10%FCS中培养。研究细胞毒性和HGF的产生，以确定AdHGF的最有效剂量。结果表明，每个细胞50 PFU的AdHGF是最有效的剂量。结果：2003年，5只狗右肝切除术后，采用聚四氟乙烯（PTFE）移植物置换IVC，植入adhgf转染的ECs。5只狗中有2只活了下来，但另外3只狗在手术后一周内死亡。死亡原因包括手术造成的无法忍受的损伤和出血。术后4周处死存活犬2只，观察肝再生及种子移植情况。有活力的内皮细胞附着的移植物上没有血栓。肝切除犬植入有籽IVC后肝脏再生与未植入有籽IVC后无明显差异。术中测定血清HGF浓度。然而，所有血清样本中的HGF水平均低于可测量值。这表明，从adhgf转染的内皮细胞在移植物上的HGF表达水平可能不足以显示生物活性。

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.