Novel approaches to cryopreservation of human pancreatic islets
Novel approaches to cryopreservation of human pancreatic islets
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DOI：
10.1097/00007890-200109270-00005
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发表时间：
2001-09-27
期刊：
影响因子：
6.2
通讯作者：
Rajotte, RV
中科院分区：
文献类型：
--
作者：
Lakey, JRT;Anderson, TJ;Rajotte, RV
背景如果要从单个人供体胰腺中回收足够的胰岛，以在分离、冷冻保存和移植后逆转1型糖尿病，则必须确定人类胰岛存活和功能的优化条件。本研究的目的是比较乙二醇（EG）与标准冷冻保护剂二甲基亚砜（DMSO）对离体人胰岛存活和功能的冷冻保护作用。此外，还研究了不同添加方案和冷冻保护剂平衡浓度的影响。通过使用胶原酶消化和不连续Ficoll梯度纯化的标准技术从人胰腺分离胰岛。新鲜分离的人胰岛的等分试样通过使用DMSO或EG冷冻保存在六组中。逐步加入冷冻保护剂以产生1.5或2.0 M的终浓度，或在一个步骤中加入至1.5 M的浓度。胰岛通过使用已建立的方案冷冻保存，并在37 ℃下培养48小时，然后评估回收率和体外活力。冷冻保存后，与标准的2.0 M DMSO（62 +/-4%）相比，在逐步方案中加入1.5 M DMSO的组中胰岛的恢复百分比（74 +/-3%，mean +/- SEM）显著更高（P
Background. Optimized conditions for survival and function of human islets must be defined if sufficient islets are to be recovered from a single human donor pancreas to reverse type-1 diabetes after isolation, cryopreservation, and transplantation. The objective of this study was to compare the cryoprotective effect of ethylene glycol (EG) with the standard cryoprotectant, dimethyl sulfoxide (DMSO), on isolated human islet survival and function. Furthermore, the effect of different addition protocols and equilibrium concentrations of the cryoprotectants were studied.Methods. Islets were isolated from human pancreata by using standard techniques of collagenase digestion and discontinuous Ficoll gradient purification. Aliquots of freshly isolated human islets were cryopreserved in six groups by using DMSO or EG. Cryoprotectants were added stepwise to produce a final concentration of 1.5 or 2.0 M, or added in a single step to a concentration of 1.5 M. Islets were cryopreserved by using established protocols and cultured for 48 hr at 37 degreesC before assessment of percentage of recovery and in vitro viability.Results. After cryopreservation, percentage of recovery of islets was significantly higher in the group treated with 1.5 M of DMSO added in a stepwise protocol (74 +/-3%, mean +/- SEM) compared with the standard 2.0 M of DMSO (62 +/-4%) (P