Ferguson J, Allsopp R H, Taylor R M, Johnston I D
Diabetologia. 1976 May;12(2):115-21. doi: 10.1007/BF00428975.
This paper reports techniques for the isolation and long term preservation of pancreatic islets from the mouse, rat and guinea pig. Islets have been isolated using a modification of a free hand microdisection procedure described by Hellerström in 1964 [1]. Isolated islets have been subjected to three preservation systems and their viability following storage assessed by light microscopy of sections stained with Gomor's aldehyde fuchsin [2] and by measuring the insulin release from islets in vitro in response to a glucose stimulus. The systems were: a) Simple cold storage in Hank's balanced salt solution at 4 degrees C. Following 15 h cold storage, histological and functional survival was 100%. This dropped to 10% at 48 h. There were no survivors following 72 h storage. b) Sub zero cell storage. In Group I (freezing rate 1 degrees C/min) histological survival was 35% and functional survival 20%. In Group II (freezing rate 5 degreees C/min with 24 h culture period after rewarming) histological survival was approximately 87% and functional survival 75%. c) Organ Culture. Islets from the guinea pig, rat and mouse showed minimal morphologic damage when cultured for 21 days in a simple organ culture system. At 28 days, histological survial was approximately 30%. Following organ culture we were unable to correlate histological and functional survival.
本文报道了从小鼠、大鼠和豚鼠中分离胰腺胰岛并进行长期保存的技术。胰岛的分离采用了对赫勒斯特伦1964年描述的徒手显微解剖程序的一种改良方法[1]。分离出的胰岛被置于三种保存系统中，并通过对用戈莫尔醛品红染色的切片进行光学显微镜观察[2]以及测量胰岛在体外对葡萄糖刺激的胰岛素释放情况，来评估储存后的活力。这些系统包括：a) 在4℃的汉克斯平衡盐溶液中简单冷藏。冷藏15小时后，组织学和功能存活率为100%。48小时后降至10%。72小时储存后无存活者。b) 零下细胞储存。在第一组（冷冻速率为1℃/分钟）中，组织学存活率为35%，功能存活率为20%。在第二组（冷冻速率为5℃/分钟，复温后有24小时培养期）中，组织学存活率约为87%，功能存活率为75%。c) 器官培养。在一个简单的器官培养系统中培养21天时，豚鼠、大鼠和小鼠的胰岛显示出最小的形态损伤。在28天时，组织学存活率约为30%。器官培养后，我们无法将组织学和功能存活率联系起来。
