Smolenski Ryszard T, Forni Monica, Maccherini Massimo, Bacci Maria Laura, Slominska Ewa M, Wang Hongjun, Fornasari Piermaria, Giovannoni Roberto, Simeone Felicetta, Zannoni Augusta, Frati Giacomo, Suzuki Ken, Yacoub Magdi H, Lavitrano Marialuisa
Heart Science Centre, Imperial College at Harefield Hospital, Harefield, Middlesex, UB9 6JH, UK.
Cardiovasc Res. 2007 Jan 1;73(1):143-52. doi: 10.1016/j.cardiores.2006.10.027. Epub 2006 Nov 6.
The use of pig hearts can solve the problem of shortage of donor hearts for transplantation. However, targeting rejection by single genetic modification was proven to be ineffective, highlighting the requirement for complex genetic modifications and more effective methods for transgenic animal production. We evaluated here whether hearts of hDAF transgenic pigs generated using our technique sperm-mediated gene transfer (SMGT) will be protected from structural damage, metabolic changes, and mechanical dysfunction during perfusion with human blood.
Hearts from control (C, n = 6) or transgenic (T, n = 5) pigs were perfused ex vivo for 4 h with fresh human blood using the ex vivo working mode system allowing monitoring of the function, metabolism, and structure.
Cardiac output (mean+/-SEM) was maintained in T constant throughout the experiment, at 3.58+/-0.36 and 3.83+/-0.14 l/min after 30 min and 4 h, respectively, while cardiac output decreased to 1.95+/-0.35 l/min in C after 30 min of perfusion (p < 0.01 vs. T). The maximum increase in coronary perfusion pressure was reduced in T to 154+/-16% as compared to C (237+/-10%, p < 0.001). Myocardial ATP after 4 h was 21.1+/-1.1 nmol/mg dry wt (similar to initial) in T, while it decreased in C to 17.2+/-1.4 (p < 0.05). Deposition of complement factors C3 and C5b9 was present in C but not in T after perfusion.
We have shown that hearts from hDAF transgenic pigs produced by SMGT are protected during perfusion with human blood and are metabolically stable and maintain mechanical function above the threshold level for life support.
使用猪心脏可解决移植供体心脏短缺的问题。然而，事实证明通过单一基因改造来靶向抑制排异反应是无效的，这凸显了进行复杂基因改造以及采用更有效方法生产转基因动物的必要性。在此，我们评估了利用我们的精子介导基因转移（SMGT）技术培育的人衰变加速因子（hDAF）转基因猪的心脏在灌注人血过程中是否能免受结构损伤、代谢变化及机械功能障碍的影响。
使用离体工作模式系统对来自对照（C组，n = 6）或转基因（T组，n = 5）猪的心脏进行4小时的新鲜人血离体灌注，该系统可监测心脏功能、代谢及结构。
在整个实验过程中，T组的心输出量（平均值±标准误）保持恒定，灌注30分钟和4小时后分别为3.58±0.36升/分钟和3.83±0.14升/分钟，而C组在灌注30分钟后心输出量降至1.95±0.35升/分钟（与T组相比，p < 0.01）。与C组相比，T组冠状动脉灌注压的最大增幅降至154±16%（C组为237±10%，p < 0.001）。4小时后，T组心肌ATP含量为21.1±1.1纳摩尔/毫克干重（与初始值相似），而C组则降至17.2±1.4（p < 0.05）。灌注后，C组出现补体因子C3和C5b9的沉积，而T组未出现。
我们已表明，通过SMGT技术培育的hDAF转基因猪的心脏在灌注人血过程中受到保护，代谢稳定，并且维持的机械功能高于生命支持的阈值水平。
