Frankel B J, Gylfe E, Hellman B, Idahl L A
J Clin Invest. 1976 Jan;57(1):47-52. doi: 10.1172/JCI108267.
Insulin content and release were measured from hand-dissected pancreatic islets from noninbred ob/ob mice after 1-5 wk storage in tissue culture medium 199 at various temperatures and glucose concentrations. After storage of islets for 1 wk at 37 degrees, 22 degrees, or 8 degrees C in 18 mM glucose medium and preincubation with 1 mM glucose, glucose-stimulated insulin release during the subsequent incubation was only 20-35% of that of fresh islets. The addition of a 4-h period at 37 degrees C with 18 mM glucose between the cold storage and perincubation restored glucose-stimulated insulin release from 8 degrees C stored islets to fresh-islet levels. Release throughout the 1-18 mM glucose range was strikingly parallel to that of fresh islets. Exposure of fresh islets to the same 4-h period increased basal release but did not affect maximal release. When islets were stored at 8 degrees C with 18 mM glucose for more than 1 wk, a short period at 37 degrees C every week was necessary for maintenance of release. After 5 wk of this procedure, glucose-stimulated insulin release was one-third that of fresh islets, or similar to that of islets stored for only 1 wk at 37 degrees C. Storage at 8 degrees C for 1 wk with 3 mM glucose, or continuously for 3 or 5 wk with 18 mM glucose, maintained islet insulin content, whereas release was lost. Thus, glucose-stimulated insulin release is best maintained by storage of pancreatic islets in tissue culture medium with a high concentration of glucose at 8 degrees C with short weekly periods at 37 degrees C.
在不同温度和葡萄糖浓度下,将非近交系ob/ob小鼠经手工解剖得到的胰岛,置于组织培养基199中储存1 - 5周后,检测其胰岛素含量和释放情况。将胰岛在18 mM葡萄糖培养基中于37℃、22℃或8℃储存1周,然后用1 mM葡萄糖预孵育,随后孵育期间葡萄糖刺激的胰岛素释放量仅为新鲜胰岛的20 - 35%。在冷藏和预孵育之间加入一段在37℃、18 mM葡萄糖条件下4小时的处理,可使8℃储存的胰岛的葡萄糖刺激的胰岛素释放恢复到新鲜胰岛水平。在1 - 18 mM葡萄糖范围内的释放情况与新鲜胰岛的情况显著平行。将新鲜胰岛暴露于相同的4小时处理会增加基础释放量,但不影响最大释放量。当胰岛在18 mM葡萄糖条件下于8℃储存超过1周时,每周需要在37℃进行短时间处理以维持释放。经过5周此程序后,葡萄糖刺激的胰岛素释放量为新鲜胰岛的三分之一,或与仅在37℃储存1周的胰岛相似。在3 mM葡萄糖条件下于8℃储存1周,或在18 mM葡萄糖条件下连续储存3或5周,可维持胰岛胰岛素含量,但释放功能丧失。因此,将胰岛储存在高浓度葡萄糖的组织培养基中,于8℃保存,并每周在37℃进行短时间处理,能最佳地维持葡萄糖刺激的胰岛素释放。