Wang An, Jank Isabel, Wei Weiwei, Schindler Claudia, Dahmen Uta
Experimental Transplantation Surgery, Department of General, Visceral and Vascular Surgery, University Hospital Jena.
Experimental Transplantation Surgery, Department of General, Visceral and Vascular Surgery, University Hospital Jena;
J Vis Exp. 2018 Oct 6(140):57991. doi: 10.3791/57991.
Organ engineering is a novel strategy to generate liver organ substitutes that can potentially be used in transplantation. Recently, in vivo liver engineering, including in vivo organ decellularization followed by repopulation, has emerged as a promising approach over ex vivo liver engineering. However, postoperative survival was not achieved. The aim of this study is to develop a novel surgical technique of in vivo selective liver lobe perfusion in rats as a prerequisite for in vivo liver engineering. We generate a circuit bypass only through the left lateral lobe. Then, the left lateral lobe is perfused with heparinized saline. The experiment is performed with 4 groups (n = 3 rats per group) based on different perfusion times of 20 min, 2 h, 3 h, and 4 h. Survival, as well as the macroscopically visible change of color and the histologically determined absence of blood cells in the portal triad and the sinusoids, is taken as an indicator for a successful model establishment. After selective perfusion of the left lateral lobe, we observe that the left lateral lobe, indeed, turned from red to faint yellow. In a histological assessment, no blood cells are visible in the branch of the portal vein, the central vein, and the sinusoids. The left lateral lobe turns red after reopening the blocked vessels. 12/12 rats survived the procedure for more than one week. We are the first to report a surgical model for in vivo single liver lobe perfusion with a long survival period of more than one week. In contrast to the previously published report, the most important advantage of the technique presented here is that perfusion of 70% of the liver is maintained throughout the whole procedure. The establishment of this technique provides a foundation for in vivo partial liver engineering in rats, including decellularization and recellularization.
器官工程是一种生成肝脏器官替代物的新策略,这些替代物有可能用于移植。最近,体内肝脏工程,包括体内器官去细胞化后再重新填充细胞,已成为一种比体外肝脏工程更有前景的方法。然而,术后存活并未实现。本研究的目的是开发一种大鼠体内选择性肝叶灌注的新手术技术,作为体内肝脏工程的前提条件。我们仅通过左外叶建立一个循环旁路。然后,用肝素化盐水灌注左外叶。根据20分钟、2小时、3小时和4小时的不同灌注时间,将实验分为4组(每组n = 3只大鼠)。将存活情况,以及肉眼可见的颜色变化和组织学确定的门静脉三联和肝血窦中无血细胞,作为成功建立模型的指标。在对左外叶进行选择性灌注后,我们观察到左外叶确实从红色变成了淡黄色。在组织学评估中,门静脉分支、中央静脉和肝血窦中均未见血细胞。重新开放阻塞血管后,左外叶恢复红色。12只大鼠全部存活超过一周。我们首次报道了一种体内单肝叶灌注的手术模型,其存活期超过一周。与之前发表的报告相比,这里介绍的技术最重要的优点是在整个过程中保持70%的肝脏灌注。该技术的建立为大鼠体内部分肝脏工程,包括去细胞化和再细胞化,奠定了基础。