Konstantinow A, Mühlbauer W, Hartinger A, von Donnersmarck G G
Department of Plastic, Reconstructive, and Hand Surgery, Bum Center, Klinikum Bogenhausen, München, Germany.
Ann Plast Surg. 1991 Jan;26(1):89-97. doi: 10.1097/00000637-199101000-00014.
A simple unit has been developed for the simultaneous passive cooling of small and large amounts of allograft or autograft split-thickness skin, as well as cultured human epidermis. An expanded polystyrene box of variable size, aluminum plates, and cellulose tissue are fused. This unit is cooled in a -70 degrees C constant-temperature mechanical refrigerator. Maximal cooling rates of -1.3 degrees C min-1 are obtained in a box with a constant wall thickness of cellulose tissue. The cooling rate can be varied by altering the number of cellulose layers. Exothermic temperature plateaus associated with skin cooled in this unit last for less than 0.3 minutes. The viability of the cryopreserved skin was determined by using up to four methods simultaneously: a dye-exclusion test with trypan blue; glucose consumption; production of lactate; and carbon-13 nuclear magnetic resonance spectroscopy. Using a cryoprotectant medium with 15% (vol/vol) glycerol for split-thickness skin 0.25 mm thick, a storage time of up to 509 days at -70 degrees C was observed, with only a small decrease in viability (trypan blue, 62.5%; glucose consumption, 71 to 90% compared with freshly harvested skin). Storage in liquid nitrogen did not significantly improve results (p greater than 0.05).
已开发出一种简单装置，用于同时对大量或少量同种异体或自体中厚皮片以及培养的人表皮进行被动冷却。一个尺寸可变的聚苯乙烯泡沫塑料盒、铝板和纤维素组织被熔合在一起。该装置在-70℃的恒温机械冰箱中冷却。在纤维素组织壁厚恒定的盒子中可获得最大冷却速率为-1.3℃/分钟。冷却速率可通过改变纤维素层数来改变。在此装置中冷却的皮肤相关的放热温度平台持续时间不到0.3分钟。通过同时使用多达四种方法来确定冷冻保存皮肤的活力：台盼蓝染料排除试验；葡萄糖消耗；乳酸生成；以及碳-13核磁共振光谱法。对于0.25毫米厚的中厚皮片，使用含15%（体积/体积）甘油的冷冻保护剂培养基，在-70℃下观察到最长储存时间为509天，活力仅有小幅下降（台盼蓝，62.5%；葡萄糖消耗，与新鲜收获的皮肤相比为71%至90%）。在液氮中储存并未显著改善结果（p＞0.05）。
