Ihm Sung-Hee, Matsumoto Ippei, Zhang Hui J, Ansite Jeffrey D, Hering Bernhard J
Department of Surgery, Diabetes Institute for Immunology and Transplantation, University of Minnesota, Minneapolis, MN, USA.
Transpl Int. 2009 Feb;22(2):207-16. doi: 10.1111/j.1432-2277.2008.00769.x. Epub 2008 Oct 13.
The Edmonton protocol for islet transplantation utilizes fresh islet grafts but other protocols increasingly transplant short-term cultured grafts mainly for practical reasons. To improve our understanding of the impact of culture pretreatment of human islets, we assessed post-transplant function by nude mouse bioassay, islet ATP, activity of stress-activated MAP kinases, and expression of stress-related genes by focused cDNA array in freshly isolated and cultured islets. Mean blood glucose levels over 4 weeks after transplantation (2000 IE) of (i) freshly isolated, (ii) cultured and preculture counted (recovery rate; 78 +/- 6%), and (iii) cultured and postculture counted islets in diabetic mice were 330 +/- 40, 277 +/- 65, and 256 +/- 52 mg/dl (i versus ii, P = 0.004; i versus iii, P = 0.002). During culture, islet ATP/DNA and ATP/ADP increased; JNK and p38 MAPK activities decreased. Among 96 genes studied, mRNA expression of heat shock protein 70 genes decreased >twofold during culture in all four pairs; expression of cyclooxygenase-2, superoxide dismutase-2, interleukin-6 and cytochromes P450 1A1 genes increased. Our results show that culturing human islets before transplantation is not disadvantageous in regard of functional recovery from changes induced by nonphysiologic stimuli during islet isolation. The increase in expression of several stress-related genes during culture also shows that improving culture conditions may further enhance post-transplant islet function.
埃德蒙顿胰岛移植方案采用新鲜胰岛移植物，但其他方案越来越多地移植短期培养的移植物，主要是出于实际原因。为了更好地理解人胰岛培养预处理的影响，我们通过裸鼠生物测定、胰岛ATP、应激激活的丝裂原活化蛋白激酶（MAPK）活性以及聚焦cDNA阵列检测新鲜分离和培养的胰岛中应激相关基因的表达，来评估移植后的功能。将（i）新鲜分离的、（ii）培养并在培养前计数（回收率；78±6%）以及（iii）培养并在培养后计数的胰岛（2000 IE）移植到糖尿病小鼠体内，移植后4周的平均血糖水平分别为330±40、277±65和256±52 mg/dl（i与ii相比，P = 0.004；i与iii相比，P = 0.002）。在培养过程中，胰岛ATP/DNA和ATP/ADP增加；JNK和p38 MAPK活性降低。在研究的96个基因中，所有四对胰岛在培养过程中热休克蛋白70基因的mRNA表达均下降两倍以上；环氧合酶-2、超氧化物歧化酶-2、白细胞介素-6和细胞色素P450 1A1基因的表达增加。我们的结果表明，移植前培养人胰岛在胰岛分离过程中非生理性刺激引起的功能恢复方面并无不利影响。培养过程中几个应激相关基因表达的增加也表明，改善培养条件可能进一步增强移植后胰岛的功能。
