Robertson G S, Chadwick D R, Davies J, Rose S, Contractor H, James R F, Bell P R, London N J
Department of Surgery, Leicester Royal Infirmary, England.
Transplantation. 1994 Feb;57(3):346-54. doi: 10.1097/00007890-199402150-00006.
The purification of human pancreatic islets before transplantation relies on the density-dependent separation of islets from exocrine fragments after collagenase digestion of the donor pancreas. The results vary among pancreases despite increasing automation of the digestion and purification processes, reflecting variations in the overlapping densities of islets and contaminating exocrine tissue. Hypothermic storage of both the pancreas and the pancreatic digest alters cell volumes and tissue densities, thereby affecting islet purification. By biochemical analysis of the isopycnic distribution of islets and exocrine tissue fragments from 23 human pancreases on linear continuous density gradients, the effect of various solutions for cold storage of pancreatic digest was studied. The use of the University of Wisconsin cold storage solution, which resulted in a significant decrease in digest volume (P = 0.006) and increase in the densities of both exocrine tissue (P = 0.001) and islets (P = 0.005), produced a significant improvement in islet purity compared with tissue culture medium (P = 0.035), predominantly due to the inclusion of a colloid, which increased the difference in density between exocrine tissue and islets. The addition of large molecular weight cellular impermeants without alteration in the concentration of permeable anions produced no effect. The results of this study support the concept that the use of solutions that minimize cell swelling throughout the process of islet purification would result in significant improvements in density-dependent islet separation, and that such solutions should contain a colloid.
人胰岛移植前的纯化依赖于在对供体胰腺进行胶原酶消化后，根据密度将胰岛与外分泌片段分离。尽管消化和纯化过程的自动化程度不断提高，但不同胰腺的结果仍存在差异，这反映了胰岛与受污染外分泌组织重叠密度的变化。胰腺及其消化物的低温保存会改变细胞体积和组织密度，从而影响胰岛纯化。通过对23个人类胰腺的胰岛和外分泌组织片段在线性连续密度梯度上的等密度分布进行生化分析，研究了各种胰腺消化物冷藏溶液的效果。与组织培养基相比，使用威斯康星大学冷藏溶液可使消化物体积显著减少（P = 0.006），外分泌组织（P = 0.001）和胰岛（P = 0.005）的密度增加，胰岛纯度得到显著提高（P = 0.035），这主要是因为加入了一种胶体，增加了外分泌组织与胰岛之间的密度差异。添加大分子细胞非渗透剂而不改变可渗透阴离子的浓度没有效果。本研究结果支持这样的概念，即在胰岛纯化过程中使用能使细胞肿胀最小化的溶液将显著改善基于密度的胰岛分离，并且这种溶液应含有一种胶体。
