Ohzato H, Gotoh M, Monden M, Dono K, Kanai T, Mori T
Department of Surgery II, Osaka University Medical School, Japan.
Transplantation. 1991 Mar;51(3):566-70. doi: 10.1097/00007890-199103000-00004.
This study tried to improve the number of viable islets isolated from a pancreas because a sufficient number cannot be obtained when the organ is preserved in the manner used for pancreas transplantation. The mechanism involved in the decrease in islet yield during preservation was studied to try to develop a better method for islet preparation. First, the integrity of the ductal system was compared between fresh and 6-hr simply preserved (in Hanks' balanced salt solution) rat pancreases. The ductal pressure after ductal injection of HBSS reached a plateau earlier and was significantly lower for the preserved pancreases (0.073 +/- 0.026 min, 410 +/- 17 mmHg, n = 5) than for the fresh ones (0.176 +/- 0.086 min, 561 +/- 103 mmHg, n = 7, P less than 0.05). Second, the extent of pancreatic distention was examined following ductal injection of barium gelatin solution. Solution leakage occurred earlier and distention was less in the preserved pancreas. In addition, the gelatin was found in the capillaries within some islets of the preserved pancreas. These results indicated that the preservation led to a rapid loss of integrity of the ductal system before collagenase injection. We therefore tested the efficacy of ductal collagenase injection at the time of harvesting: 15 ml of 1.0 mg/ml collagenase HBSS was intraductally injected and the pancreas was preserved at 4 degrees C for 2, 4, 6, and 24 hr. The isolation procedure was similar to that used for the fresh pancreas. The yield was significantly better than that of the simply preserved pancreas at 4 hr (241 +/- 22, n = 3, vs. 140 +/- 58, n = 3, P less than 0.05) and at 6 hr (171 +/- 58, n = 14, vs. 32 +/- 33, n = 6, P less than 0.01). These isolated islets were spherical-oval and their viability was confirmed by the ability to reverse STZ-induced diabetes in mice. These results indicated that the integrity of the ductal system, which is necessary for distention of the whole pancreas, was lost during preservation. To solve this problem, ductal collagenase injection should be done at the time of pancreas harvesting and then followed by simple preservation. This method is recommended to obtain viable islets from a preserved pancreas.
本研究试图提高从胰腺分离出的活胰岛数量，因为当按照用于胰腺移植的方式保存器官时，无法获得足够数量的胰岛。研究了保存过程中胰岛产量下降所涉及的机制，以试图开发一种更好的胰岛制备方法。首先，比较了新鲜大鼠胰腺和简单保存6小时（在汉克斯平衡盐溶液中）的大鼠胰腺的导管系统完整性。向导管内注射HBSS后的导管压力达到平稳期的时间更早，并且保存的胰腺（0.073±0.026分钟，410±17 mmHg，n = 5）显著低于新鲜胰腺（0.176±0.086分钟，561±103 mmHg，n = 7，P＜0.05）。其次，在向导管内注射钡明胶溶液后检查胰腺扩张程度。保存的胰腺中溶液渗漏更早出现且扩张程度更小。此外，在保存的胰腺的一些胰岛内的毛细血管中发现了明胶。这些结果表明，在注射胶原酶之前，保存导致导管系统完整性迅速丧失。因此，我们测试了在收获时向导管内注射胶原酶的效果：向导管内注射15 ml 1.0 mg/ml胶原酶HBSS，并将胰腺在4℃保存2、4、6和24小时。分离程序与用于新鲜胰腺的程序相似。在4小时（241±22，n = 3，对比140±58，n = 3，P＜0.05）和6小时（171±58，n = 14，对比32±33，n = 6，P＜0.01）时，产量显著高于简单保存的胰腺。这些分离出的胰岛呈球形 - 椭圆形，并且它们的活力通过逆转小鼠链脲佐菌素诱导的糖尿病的能力得以证实。这些结果表明，对于整个胰腺扩张所必需的导管系统完整性在保存过程中丧失了。为了解决这个问题，应在胰腺收获时进行导管内胶原酶注射，然后进行简单保存。推荐使用这种方法从保存的胰腺中获得活胰岛。
