Kim Hyoung-Il, Yu Jae Eun, Lee Song Yi, Sul A Young, Jang Min Seok, Rashid M A, Park Sang Gyu, Kim Sang Jun, Park Chung-Gyu, Kim Jae Hyeon, Park Kyong Soo
Xenotransplantation Research Center, Seoul 110-744, Republic of Korea.
Cell Transplant. 2009;18(1):31-7. doi: 10.3727/096368909788237113.
Instant blood-mediated inflammatory reaction (IBMIR) causes rapid islet loss in portal vein islet transplantation. Endothelial cells are known to protect against complement-mediated lysis and activation of coagulation. We tested composite pig islet-human endothelial cell grafts as a strategy to overcome IBMIR. Porcine islets were cocultured with human endothelial cells in specially modified culture medium composed of M199 and M200 for 1-9 days. A positive control group, negative control group, and the endothelial cell-coated group were examined with an in vitro tubing loop assay using human blood. The endothelial cell-coated group was subdivided and analyzed by degree of surface coverage by endothelial cells (< or = 50% vs. > 50%) or coculture time (< 5 days vs. > or = 5 days). Platelet consumption and complement and coagulation activation were assessed by platelet count, C3a, and thrombin-antithrombin complex (TAT), respectively. After 60-min incubation in human blood, the endothelial cell-coated group showed platelet consumption inhibition and low C3a and TAT assay results compared to uncoated controls. When the endothelial cell-coated group was subdivided by degree of surface coverage, the < or = 50% coated group showed less platelet consumption and less activation of complement and coagulation compared with the positive control (uncoated) group. On analysis by coculture time, only the subgroup cocultured for < 5 days showed the same protective effect. Human endothelial cell-coated pig islets, especially the partially coated and short-term cocultured pig islet-human endothelial cell composites, reduced all components of IBMIR. If the optimal endothelial cell-islet coculture method could be identified, human endothelial cell coating of pig islets would offer new strategies to improve xenogenic islet transplantation outcomes.
即时血液介导的炎症反应（IBMIR）导致门静脉胰岛移植中胰岛迅速丢失。已知内皮细胞可防止补体介导的溶解和凝血激活。我们测试了复合猪胰岛-人内皮细胞移植物作为克服IBMIR的一种策略。将猪胰岛与人内皮细胞在由M199和M200组成的特殊改良培养基中共培养1 - 9天。使用人血通过体外管路环试验对阳性对照组、阴性对照组和内皮细胞包被组进行检测。内皮细胞包被组根据内皮细胞表面覆盖程度（≤50%与>50%）或共培养时间（<5天与≥5天）进行细分和分析。分别通过血小板计数、C3a和凝血酶 - 抗凝血酶复合物（TAT）评估血小板消耗以及补体和凝血激活情况。在人血中孵育60分钟后，与未包被的对照组相比，内皮细胞包被组显示出血小板消耗受到抑制，且C3a和TAT检测结果较低。当根据表面覆盖程度对内皮细胞包被组进行细分时，≤50%包被组与阳性对照（未包被）组相比，血小板消耗更少，补体和凝血激活也更少。根据共培养时间进行分析时，只有共培养<5天的亚组显示出相同的保护作用。人内皮细胞包被的猪胰岛，特别是部分包被和短期共培养的猪胰岛 - 人内皮细胞复合物，减少了IBMIR的所有成分。如果能够确定最佳的内皮细胞 - 胰岛共培养方法，猪胰岛的人内皮细胞包被将为改善异种胰岛移植结果提供新的策略。
