Balamurugan A N, Chang Yigang, Fung John J, Trucco Massimo, Bottino Rita
Department of Surgery, Thomas E. Starzl Transplantation Institute, University of Pittsburgh, School of Medicine, Pittsburgh, PA, USA.
Am J Transplant. 2003 Sep;3(9):1135-42. doi: 10.1046/j.1600-6143.2003.00184.x.
Worldwide growing interest in reproducing the result of the Edmonton protocol in islet transplantation trials poses the problem of paucity of donors to supply sufficient amount of islets for clinical use. Improved outcomes include finding better ways to obtain higher yields from every donor organ processed and the possibility of extending islet isolation processing to glands of suboptimal quality. In order to optimize enzymatic digestion of marginal donor organs, we have modified the technique of tissue collection following enzymatic digestion of human pancreatic organs, allowing for reduced time of exposure of free islets to warm Liberase trade mark solution. Our results indicate that better controlled exposure to enzyme yields: (i) higher islet numbers; (ii) complete dissociation of all parts of pancreatic tissue; (iii) successful islet harvest from organs otherwise excluded. We also show that by limiting the exposure of free islets to enzyme solution, islet fragmentation and loss of insulin content are reduced. We further support evidence that enzymatic digestion may contribute to impairment of insulin secretory capacity of the islets in vitro during culture.
全球范围内，胰岛移植试验对重现埃德蒙顿方案结果的兴趣日益浓厚，这带来了供体短缺的问题，难以提供足够数量的胰岛用于临床。改善结果包括找到更好的方法，从每个处理过的供体器官中获得更高的产量，以及将胰岛分离处理扩展到质量欠佳腺体的可能性。为了优化对边缘供体器官的酶消化，我们改进了人胰腺器官酶消化后的组织收集技术，减少了游离胰岛暴露于温热的 Liberase 商标溶液中的时间。我们的结果表明，对酶的更精确控制的暴露可产生：（i）更高的胰岛数量；（ii）胰腺组织所有部分的完全解离；（iii）从原本被排除的器官中成功收获胰岛。我们还表明，通过限制游离胰岛暴露于酶溶液的时间，可减少胰岛破碎和胰岛素含量的损失。我们进一步支持了这样的证据，即酶消化可能在培养过程中导致体外胰岛胰岛素分泌能力受损。
