Department of Surgery, Campus Charité Mitte | Campus Virchow-Klinikum, Experimental Surgery, Charité-Universitätsmedizin Berlin, Corporate Member of Freie Universität Berlin, Humboldt-Universität zu Berlin, and Berlin Institute of Health, Berlin, Germany.
Department of Pathology, Charité-Universitätsmedizin Berlin, Corporate Member of Freie Universität Berlin, Humboldt-Universität zu Berlin, and Berlin Institute of Health, Berlin, Germany.
Tissue Eng Part A. 2020 Jan;26(1-2):57-65. doi: 10.1089/ten.TEA.2019.0152. Epub 2019 Sep 18.
liver machine perfusion (MP) is a promising alternative for preservation of liver grafts from extended criteria donors. Small animal models can be used to evaluate different perfusion conditions. We here describe the development of a miniaturized MP system for rat liver grafts, evaluating cell-free and erythrocyte-based perfusion solutions, subnormothermic and normothermic temperatures, and dialysis. A perfusion chamber was designed after a suitable liver position was identified. Normothermic liver perfusion (NEVLP) required supplementation of erythrocytes to reduce cell damage. Perfusion with erythrocytes led to rising potassium levels after 12 h (NEVLP, 16.2 mM, interquartile range [IQR]: 5.7 and subnormothermic liver perfusion [SNEVLP], 12.8 mM, IQR: 3.5), which were normalized by dialysis using a laboratory dialysis membrane (NEVLP, 6.2 mM, IQR: 0.5 and SNEVLP, 5.3 mM, IQR: 0.1; = 0.004). Livers treated with NEVLP conditions showed higher bile production (18.52 mg/h/g, IQR: 8.2) compared to livers perfused under SNEVLP conditions (0.4 mg/h/g, IQR: 1.2, = 0.01). Reducing the perfusion volume from 100 to 50 mL allowed for higher erythrocyte concentrations, leading to significantly lower transaminases (15.75 U/L/mL, IQR: 2.29 vs. 5.97 U/L/mL, IQR: 18.07, = 0.002). In conclusion, a well-designed perfusion system, appropriate oxygen carriers, dialysis, and miniaturization of the perfusion volume are critical features for successful miniaturized liver MP. Impact Statement liver machine perfusion (MP) is an emerging preservation alternative to static cold storage. Even though clinical studies have shown benefits for extended criteria donor grafts, standardized systems for perfusion of rat liver grafts are not available, which are inevitable for large-scaled studies on liver reconditioning by MP. We here comprehensively describe the development of an rat liver perfusion system that can be used as modular setting in various approaches of liver MP. We describe pitfalls and techniques that might be of interest when establishing such perfusion systems for basic and translational research.
肝脏机器灌注（MP）是一种有前途的替代方法，可以保存来自扩展标准供体的肝移植物。小动物模型可用于评估不同的灌注条件。我们在这里描述了一种用于大鼠肝移植物的小型化 MP 系统的开发，评估了无细胞和基于红细胞的灌注溶液、亚体温和体温以及透析。在确定合适的肝脏位置后，设计了一个灌注室。体温 MP（NEVLP）需要补充红细胞以减少细胞损伤。用红细胞灌注 12 小时后，钾水平升高（NEVLP，16.2mM，四分位距[IQR]：5.7 和亚体温 MP[SNEVLP]，12.8mM，IQR：3.5），使用实验室透析膜进行透析可使钾水平正常化（NEVLP，6.2mM，IQR：0.5 和 SNEVLP，5.3mM，IQR：0.1； = 0.004）。用 NEVLP 处理的肝脏显示出比在 SNEVLP 条件下灌注的肝脏更高的胆汁产生（18.52mg/h/g，IQR：8.2）（=0.01）。将灌注体积从 100 减少到 50 mL 允许更高的红细胞浓度，导致转氨酶显著降低（15.75U/L/mL，IQR：2.29 与 5.97U/L/mL，IQR：18.07， = 0.002）。总之，设计良好的灌注系统、合适的氧载体、透析和灌注体积的小型化是成功的小型化肝脏 MP 的关键特征。 影响陈述 肝脏机器灌注（MP）是静态冷藏的一种新兴替代保存方法。尽管临床研究表明，对于扩展标准供体移植物有好处，但大鼠肝移植物灌注的标准化系统尚不可用，这对于通过 MP 对肝再灌注进行大规模研究是必不可少的。我们在这里全面描述了一种大鼠肝脏灌注系统的开发，该系统可作为各种肝脏 MP 方法的模块化设置。我们描述了在为基础和转化研究建立此类灌注系统时可能感兴趣的缺陷和技术。
