University of Michigan, Ann Arbor; University of Montreal;
University of Michigan, Ann Arbor.
J Vis Exp. 2022 Dec 23(190). doi: 10.3791/64384.
Primary graft dysfunction (PGD) remains the leading cause of early death following heart transplantation. Prolonged ischemic time during cold preservation is an important risk factor for PGD, and reliable evaluation of cardiac function is essential to study the functional responses of the donor heart after cold preservation. The accompanying video describes a technique to assess murine right and left ventricular function using ex vivo perfusion based in a Langendorff model after cold preservation for different durations. In brief, the heart is isolated and stored in a cold histidine-tryptophan-ketoglutarate (HTK) solution. Then, the heart is perfused with a Kreb buffer in a Langendorff model for 60 min. A silicone balloon is inserted into the left and right ventricle, and cardiac functional parameters are recorded (dP/dt, pressure-volume relationships). This protocol allows the reliable evaluation of cardiac function after different heart preservation protocols. Importantly, this technique allows the study of cardiac preservation responses specifically in native cardiac cells. The use of very small murine hearts allows access to an enormous array of transgenic mice to investigate the mechanisms of PGD.
原发性移植心脏功能障碍(PGD)仍然是心脏移植后早期死亡的主要原因。冷保存期间的缺血时间延长是 PGD 的一个重要危险因素,因此可靠地评估供心的功能对于研究冷保存后供心的功能反应至关重要。本视频描述了一种在冷保存不同时间后使用基于 Langendorff 模型的体外灌流来评估小鼠右心室和左心室功能的技术。简而言之,心脏被分离出来并储存在含有组氨酸-色氨酸-酮戊二酸(HTK)的冷溶液中。然后,心脏在 Langendorff 模型中用 Krebs 缓冲液灌流 60 分钟。将一个硅酮球囊插入左心室和右心室,并记录心脏功能参数(dp/dt,压力-容积关系)。该方案可用于评估不同心脏保存方案后的心脏功能。重要的是,该技术允许专门研究天然心脏细胞中的心脏保存反应。使用非常小的小鼠心脏,可以访问大量的转基因小鼠,以研究 PGD 的机制。