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Selective Osmotic Shock for Islet Isolation in the Cadaveric Canine Pancreas

机译:选择性渗透压休克法在犬尸胰腺胰岛分离中的应用

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摘要

Currently, islet isolation is performed using harsh collagenases that cause nonspecific injury to both islets and exocrine tissue, negatively affecting the outcome of cell transplantation. We evaluated a novel islet isolation protocol utilizing high concentrations of glucose to cause selective osmotic shock (SOS). Islets have a membrane glucose transporter that allows adaptation to changes in glucose concentrations while exocrine tissue can be selectively destroyed by these osmolar shifts. Canine pancreata were obtained within 15 min after euthanasia from animals (n = 6) euthanized for reasons unrelated to this study. Each pancreas was divided into 4 segments that were randomized to receive 300 mOsm glucose for 20 min (group 1), 600 mOsm for 20 min (group 2), 300 mOsm for 40 min (group 3), or 600 mOsm for 40 min (group 4). Islet yield, purity, and viability were compared between groups. Mean ± standard error of the mean islet yield for groups 1 to 4 was 428 ± 159, 560 ± 257, 878 ± 443, and 990 ± 394 islet equivalents per gram, respectively. Purity ranged from 37% to 45% without the use of density gradient centrifugation and was not significantly different between groups. Islet cell viability was excellent overall (89%) and did not differ between treatment protocol. Islet function was best in groups treated with 300 mOsm of glucose (stimulation index [SI] = 3.3), suggesting that the lower concentration of glucose may be preferred for use in canine islet isolation. SOS provides a widely available means for researchers to isolate canine islets for use in islet transplantation or in studies of canine islet physiology.
机译:当前,使用苛刻的胶原酶进行胰岛分离,所述胶原酶对胰岛和外分泌组织均造成非特异性损伤,对细胞移植的结果产生负面影响。我们评估了利用高浓度葡萄糖引起选择性渗透休克(SOS)的新型胰岛分离方案。胰岛具有膜葡萄糖转运蛋白,它可以适应葡萄糖浓度的变化,而外分泌组织可以被这些渗透压变化选择性破坏。安乐死后15分钟内,从与本研究无关的原因中对动物(n = 6)实施安乐死获得了犬胰脏。将每个胰腺分成4个部分,随机分为300分钟,300分钟,300分钟,600分钟(3组)或600分钟(40分钟)(第1组),300分钟(300组),20分钟(第2组),600毫秒(4组)。组4)。比较两组之间的胰岛产量,纯度和生存力。第1至第4组的平均胰岛产量的平均±标准误差分别为每克428±159、560±257、878±443和990±394胰岛当量。在不使用密度梯度离心的情况下,纯度范围为37%至45%,各组之间的差异不显着。胰岛细胞生存力总体上非常出色(89%),并且在治疗方案之间没有差异。在用300 mOsm葡萄糖治疗的组中,胰岛功能最好(刺激指数[SI] = 3.3),这表明较低的葡萄糖浓度可能更适合用于犬胰岛分离。 SOS为研究人员分离犬胰岛提供了广泛可用的手段,可用于胰岛移植或犬胰岛生理学研究。

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