Wang T C, Connery C P, Batty P R, Hicks G L, DeWeese J A, Layne J R
Department of Surgery, University of Rochester, New York 14642.
Cryobiology. 1991 Apr;28(2):171-6. doi: 10.1016/0011-2240(91)90019-k.
The present study adapted the overwintering strategy employed by freeze-tolerant amphibians and reptiles to freeze-preserve the isolated rat heart. The heart was flushed with a cardioplegic solution and supercooled to -1.2 and -3 degrees C. Then freezing was induced by inoculation of ice crystal. The viability of the heart explant was assessed after reanimation by the isolated working heart perfusion. There was no recovery of function in hearts flushed with solution containing 0.28 mM CaCl2. Lowering the concentration of CaCl2 to 0.15 mM, however, rendered good functional return. Furthermore, inclusion of 50 mM glycerol in the flush solution dramatically improved functional preservation. Under the best conditions defined here, the recoveries of aortic flow, coronary flow, cardiac output, systolic pressure, and work in hearts stored at -1.2 degrees C for 3 h were 72.8 +/- 6.8, 87.2 +/- 4.2, 77.6 +/- 5.4, 83.4 +/- 2.8, and 66.6 +/- 5.9% (mean +/- SEM, n = 8) of the unstored control levels, respectively. The myocardial ice content was 18.6 +/- 5.4% (n = 5) of tissue water. Prolonging the storage time to 5 h increased the ice content to 45.3 +/- 8.1% and reduced the recovery of cardiac output to 23 +/- 11% of the control value (mean +/- SEM, n = 5). Hearts frozen at -3 degrees C for 1.5 h showed 29.4 +/- 8.7% (n = 3) of control cardiac output during reperfusion. This novel approach may provide an opportunity to advance our knowledge about freezing preservation of not only the heart but other solid organs as well.
本研究采用了耐冻两栖动物和爬行动物的越冬策略来冷冻保存离体大鼠心脏。用心脏停搏液冲洗心脏，并将其过冷至-1.2℃和-3℃。然后通过接种冰晶诱导冷冻。复苏后通过离体工作心脏灌注评估心脏外植体的活力。用含0.28 mM氯化钙的溶液冲洗的心脏没有功能恢复。然而，将氯化钙浓度降至0.15 mM可实现良好的功能恢复。此外，在冲洗液中加入50 mM甘油可显著改善功能保存。在此处定义的最佳条件下，在-1.2℃储存3小时的心脏中，主动脉流量、冠状动脉流量、心输出量、收缩压和功的恢复分别为未储存对照水平的72.8±6.8%、87.2±4.2%、77.6±5.4%、83.4±2.8%和66.6±5.9%（平均值±标准误，n = 8）。心肌冰含量为组织水的18.6±5.4%（n = 5）。将储存时间延长至5小时会使冰含量增加至45.3±8.1%，并使心输出量的恢复降低至对照值的23±11%（平均值±标准误，n = 5）。在-3℃冷冻1.5小时的心脏在再灌注期间的心输出量为对照的29.4±8.7%（n = 3）。这种新方法可能为增进我们对不仅心脏而且其他实体器官冷冻保存的了解提供机会。
