使用羟乙基淀粉和二甲亚砜作为冷冻保护剂的胰岛冷冻保存简化方法。

Simplified method for cryopreservation of islets using hydroxyethyl starch and dimethyl sulfoxide as cryoprotectants.

作者信息

Maruyama M, Kenmochi T, Sakamoto K, Arita S, Iwashita C, Kashiwabara H

机构信息

Department of Surgery, Sakura National Hospital, Chiba, Japan.

出版信息

Transplant Proc. 2004 May;36(4):1133-4. doi: 10.1016/j.transproceed.2004.04.016.

DOI:10.1016/j.transproceed.2004.04.016

PMID:15194395

Abstract

Cryopreservation is an ideal method for long-term storage of human islets. Dimethyl sulfoxide (DMSO) has been used as an intracellular cryoprotectant. However, because of its toxicity, DMSO has to be added stepwise and diluted stepwise with sucrose. We combined hydroxyethyl starch (HES) as an extracellular cryoprotectant with DMSO to simplify the freeze-thawing procedure. Islets were isolated from the pancreas of beagle dogs by an automated digestion method and Ficoll purification. After overnight culture, the islets were cryogeneically stored using cooling by a programmed freezing system. After 4-week storage in liquid nitrogen, the container was rapidly thawed in a 37 degrees C water bath. The function of the islets was assessed upon static incubation immediately after thawing, showing a recovery rate of 71.16% +/- 20.14% and a stimulation index of 1.80 +/- 0.78. In conclusion use of HES allowed a decrease in DMSO concentration and simplified the freeze-thawing procedure for islets.

摘要

冷冻保存是长期储存人胰岛的理想方法。二甲基亚砜（DMSO）一直被用作细胞内冷冻保护剂。然而，由于其毒性，DMSO必须逐步添加并用蔗糖逐步稀释。我们将羟乙基淀粉（HES）作为细胞外冷冻保护剂与DMSO联合使用，以简化冻融程序。通过自动消化法和Ficoll纯化从比格犬的胰腺中分离胰岛。过夜培养后，使用程序冷冻系统冷却将胰岛进行低温保存。在液氮中储存4周后，将容器在37℃水浴中快速解冻。解冻后立即进行静态孵育评估胰岛功能，显示回收率为71.16%±20.14%，刺激指数为1.80±0.78。总之，使用HES可降低DMSO浓度并简化胰岛的冻融程序。