Tsujimura Toshiaki, Kuroda Yoshikazu, Churchill Thomas A, Avila Jose G, Kin Tatsuya, Shapiro A M James, Lakey Jonathan R T
Surgical-Medical Research Institute, Department of Surgery, University of Alberta, Edmonton, Alberta T6G2N8, Canada.
Cell Transplant. 2004;13(1):67-73. doi: 10.3727/000000004772664914.
A two-layer cold storage method (TLM) allows sufficient oxygen delivery to pancreata during preservation and resuscitates the viability of ischemically damaged pancreata in the canine pancreas transplant model. In this study, we applied a short-term preservation of the TLM to human pancreata after prolonged cold ischemia prior to islet isolation, and investigated the mechanisms of resuscitation of the ischemically damaged human pancreas by the TLM. Human pancreata were procured from cadaveric donors and preserved by the TLM for 3.2 +/- 0.5 h after 11.1 +/- 0.9 h of cold storage in UW (TLM group), or by cold UW alone for 11.0 +/- 0.3 h (UW group). Islet isolations of all pancreata were performed using the Edmonton protocol. Islet recovery and in vitro functional viability of isolated islets were significantly increased in the TLM group compared with the UW group. According to the criteria of the Edmonton protocol, 10/14 cases (71%) in the TLM group were transplanted to patients with type I diabetes mellitus compared with only 5/21 cases (24%) in the UW group. In the metabolic assessment of human pancreata, levels of energetic parameters (ATP, total adenylates, and energy charge) were significantly increased, and malondialdehyde (MDA) levels were significantly decreased after the TLM preservation. There was no observable change in the incidence or degree of mitochondrial injury after the TLM preservation. Additional short-term storage by the TLM resuscitates the ischemically damaged human pancreas by regenerating the energetic status and prevents further damage by oxidative stress, ultimately leading to improvements of islet recovery and in vitro function. Use of the TLM following prolonged storage in UW provides an excellent adjunctive protocol for treating human pancreata for the rigors of the islet isolation process.
一种双层冷藏方法（TLM）可在保存过程中为胰腺提供充足的氧气，并在犬胰腺移植模型中恢复缺血损伤胰腺的活力。在本研究中，我们在胰岛分离前对长时间冷缺血后的人胰腺应用TLM进行短期保存，并研究TLM恢复缺血损伤人胰腺活力的机制。人胰腺取自尸体供体，在UW中冷藏11.1±0.9小时后，采用TLM保存3.2±0.5小时（TLM组），或仅用冷UW保存11.0±0.3小时（UW组）。所有胰腺的胰岛分离均采用埃德蒙顿方案进行。与UW组相比，TLM组分离胰岛的回收率和体外功能活力显著提高。根据埃德蒙顿方案的标准，TLM组14例中有10例（71%）移植给了I型糖尿病患者，而UW组21例中只有5例（24%）。在对人胰腺的代谢评估中，TLM保存后能量参数（ATP、总腺苷酸和能量电荷）水平显著升高，丙二醛（MDA）水平显著降低。TLM保存后线粒体损伤的发生率或程度没有明显变化。通过TLM进行额外的短期保存可通过恢复能量状态来恢复缺血损伤的人胰腺，并防止氧化应激进一步损伤，最终导致胰岛回收率和体外功能的改善。在UW中长时间保存后使用TLM为处理人胰腺以应对胰岛分离过程的严苛要求提供了一种出色的辅助方案。
