Cryopreservation of human pancreatic islets from non-heart-beating donors using hydroxyethyl starch and dimethyl sulfoxide as cryoprotectants
Cryopreservation of human pancreatic islets from non-heart-beating donors using hydroxyethyl starch and dimethyl sulfoxide as cryoprotectants
复制标题
DOI：
10.3727/000000008783907026
复制
发表时间：
2008-01-01
影响因子：
3.3
通讯作者：
Miyazaki, Mariko
中科院分区：
文献类型：
--
作者：
Kenmochi, Takashi;Asano, Takehide;Miyazaki, Mariko
尽管广泛使用，DMSO 对胰岛有毒。我们将羟乙基淀粉（HES）与DMSO结合，简化了冻融过程，并降低了DMSO的毒性。使用比格犬的胰岛进行了临床前研究。储存4周后，将胰岛解冻并进行检查。解冻后胰岛结构保持良好。尽管胰岛数量减少至71.2+/-20.1%，但通过解冻后静态孵育评估胰岛的功能，并显示出1.80+/-0.78的刺激指数。我们引入了这种技术来冷冻保存来自无心跳捐赠者的人类胰岛。进行了十二例人胰岛冷冻保存。将每个人体冷冻保存的样本管解冻，以评估形态、污染和内分泌功能。尽管在五个样品（41.6%）中观察到碎片，但通过显微镜和电子显微镜研究评估，其他七个（58.4%）显示出正常结构。解冻后静态孵育的刺激指数（SI）从3.37+/-3.02恶化至1.34+/-0.28。我们将解冻的胰岛分为两组：组 1 (n = 8)，SI > 1.2；组 1 (n = 8)，SI > 1.2；第 2 组 (n = 4)，SI < 1.2。第一组胰岛的正常结构率 (87%) 高于第二组 (25%)。此外，第1组冻存前的SI为4.01±3.57，高于第2组的2.11±0.72的SI。基于大动物模型临床前研究的良好结果，将该方法引入临床应用。即使来自无心跳供体的胰腺，也能成功实现胰岛冷冻保存。但功能较差的离体胰岛不宜冷冻保存用于移植。
Although widely used, DMSO is toxic for pancreatic islets. We combined hydroxyethyl starch (HES) with DMSO to simplify the procedure of freezing and thawing, and to decrease the toxicity of DMSO. A preclinical study was performed using islets from beagle dogs. After storage for 4 weeks, the islets were thawed and examined. The islet structure was well maintained after thawing. Although the number of the islets decreased to 71.2 +/- 20.1 %, the function of the islets was evaluated by static incubation after thawing and showed a 1.80 +/- 0.78 stimulation index. We have introduced this technique for the cryopreservation of human islets from non-heart-beating donors. Twelve cases of human islet cryopreservation were performed. The sample tube of each human cryopreservation was thawed to evaluate the morphology, contamination, and endocrine function. Although fragmentation was observed in five samples (41.6%), the other seven (58.4%) showed a normal structure when evaluated by microscopic and electron microscopic study. The stimulation index (SI) of static incubation deteriorated from 3.37 +/- 3.02 to 1.34 +/- 0.28 after thawing. We divided the thawed islets into two groups: group 1 (n = 8), SI > 1.2; group 2 (n = 4), SI < 1.2. The group I islets showed a higher rate of normal structure (87%) than did group 2 (25%). Moreover, the SI before cryopreservation was 4.01 +/- 3.57 in group 1, which was higher than the SI of 2.11 +/- 0.72 in group 2. Based on the good results from the preclinical study using a large-animal model, this method was introduced for clinical application. Even from the pancreata of non-heart-beating donors, a successful islet cryopreservation was achieved. However, the isolated islets with poor function should not be cryopreserved for transplantation.