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首页> 外文期刊>Current Science: A Fortnightly Journal of Research >Selective loss of exocrine tissue during islet storage at - 70 degrees C
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Selective loss of exocrine tissue during islet storage at - 70 degrees C

机译：在-70摄氏度的胰岛储存过程中外分泌组织的选择性损失

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Since two and half decades it has been shown that frozen-thawed-transplanted islets were able to normalize blood sugar in streptozotocin-diabetic rats. In experiments with adult rat islets, a number of different freezing protocols have been used, with viability demonstrated both in vitro and in vivo. Effective islet transplant programme promotion requires different forms of storage, such as cold storage of the pancreas during transport to a centre for islet isolation; tissue culture to reduce tissue immunogenicity and cryopreservation for long-term storage. It is also mandatory to have pure islet preparation without exocrine cell contamination for islet transplantation into a diabetic recipient. In our ongoing project on islet isolation and cryopreservation we came across an interesting feature on differential susceptibility and sensitivity of pancreatic cell populations to cryo storage at -70 deg C. Islets were isolated from Balb/C mice of either sex following the protocol developed by us and the resuiting islets along with acinar cells were cryopreserved at -70 deg C (Deep Freezer, SO LOW, Cincinatti, Ohio, USA). After one month of storage at -70 deg C, islet preparations were rapidly thawed at 3T'C and revived as described earliers. Revived islets were stained for their viability and specificity employing trypan blue dye exclusion test and dithizone staining, respectively. It was observed that islets showed 92 +- 2.5 percent viability and remained unstained with trypan blue, whereas the acinar cells present in the preparation showed almost zero viability, turning blue in colour. The viability of frozenthawed islets was comparable to that of freshly isolated islets (94 + 3.1) The viable islets stained positive (brick-red in colour) with dithizone indicating their specificity and identity, whereas acinar cells remained unstained with dithizone.
机译：自从两个半月以来，已经证明冷冻融化的胰岛能够使链脲佐菌素-糖尿病大鼠的血糖正常化。在成年大鼠胰岛的实验中，已经使用了许多不同的冷冻方案，并在体外和体内都证明了其可行性。有效的胰岛移植计划推广需要不同形式的存储，例如在运输到胰岛隔离中心的过程中胰腺的冷藏;组织培养，以减少组织的免疫原性和长期保存的低温保存。对于将胰岛移植到糖尿病受体中的胰岛，也必须进行纯净的胰岛制备，以免受到外分泌细胞的污染。在我们正在进行的有关胰岛分离和冷冻保存的项目中，我们遇到了一个有趣的功能，涉及胰腺细胞群体对-70℃冷冻保存的差异敏感性和敏感性。按照我们制定的方案，从任何性别的Balb / C小鼠中分离出胰岛并将胰岛细胞和腺泡细胞在-70℃冷冻保存（Deep Freezer，SO LOW，辛辛那提，俄亥俄州，美国）。在-70摄氏度下保存一个月后，胰岛制剂在3T'C迅速解冻，并如前所述恢复了活力。分别用台盼蓝染料排斥试验和双硫zone染色法对再生的胰岛的生存力和特异性进行染色。观察到胰岛显示出92±2.5％的生存力，并没有被锥虫蓝染色，而制剂中存在的腺泡细胞则显示出几乎为零的生存力，变成了蓝色。冷冻融化的胰岛的活力与新鲜分离的胰岛的活力相当（94 + 3.1）。活的胰岛被双硫zone染色呈阳性（砖红色），表明它们的特异性和同一性，而腺泡细胞未被双硫zone染色。

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