Zacharovova K, Berkova Z, Spacek T, Kriz J, Dovolilova E, Girman P, Koblas T, Jezek P, Saudek F
Laboratory of Pancreatic Islets, Institute for Clinical and Experimental Medicine, Videnska 1958/9, 140-21 Prague, Czech Republic.
Transplant Proc. 2005 Oct;37(8):3454-6. doi: 10.1016/j.transproceed.2005.09.053.
In order to assess the quality of freshly isolated and cultivated pancreatic islets designed for experimental transplantation in rats we combined the vitality staining test, in vitro measurement of insulin secretion capacity, and assessment of islet respiration. Oxygen consumption was measured using the respirometer Oxygraph 2K equipped with polarographic oxygen sensors. The results of oxymetry demonstrated a linear correlation between islet number and oxygen consumption. Respiration per unit of viable islet tissue was constant. Oxygen consumption tests were in good correlation with the results of insulin release assays, with a correlation coefficient of 0.82. We found no significant differences in all three vitality-testing methods performed with fresh and 24-hour cultivated islets (P > .05). We conclude that polarographic oxymetry provides a fast and easy evaluation test of islet quality. After appropriate standardization, the oxymetric technique can be used for routine clinical pretransplant islet quality testing. In addition, cell membrane integrity and mitochondrial function could be assessed after addition of specific respiration inhibitors or stimulators.
为评估用于大鼠实验性移植的新鲜分离和培养的胰岛质量,我们结合了活力染色试验、胰岛素分泌能力的体外测量以及胰岛呼吸评估。使用配备极谱氧传感器的呼吸计Oxygraph 2K测量氧气消耗。血氧测定结果表明胰岛数量与氧气消耗之间存在线性相关。每单位存活胰岛组织的呼吸是恒定的。氧气消耗测试与胰岛素释放试验结果具有良好的相关性,相关系数为0.82。我们发现,对新鲜和培养24小时的胰岛进行的所有三种活力测试方法均无显著差异(P>0.05)。我们得出结论,极谱血氧测定法提供了一种快速简便的胰岛质量评估测试。经过适当标准化后,血氧测定技术可用于常规临床移植前胰岛质量检测。此外,加入特定的呼吸抑制剂或刺激剂后,可以评估细胞膜完整性和线粒体功能。