Yamanaka Takahiro, Goto Teppei, Hirabayashi Masumi, Hochi Shinichi
1 Department of Textile Science and Technology, Graduate School of Science and Technology, Shinshu University , Ueda, Japan .
2 Center for Genetic Analysis of Behavior, National Institute for Physiological Sciences , Okazaki, Japan .
Biopreserv Biobank. 2017 Oct;15(5):457-462. doi: 10.1089/bio.2017.0044. Epub 2017 Sep 5.
The practical requirements of islet transplantation necessitate that a large quantity of pancreatic islets be cryopreserved for a long period of time in a simple and convenient manner. We cryopreserved rat islets (size range 101-150 μm in mean diameter) by vitrification with either a Cryotop device or a ø = 57-μm nylon mesh device in units of 10 islets, or by conventional freezing with a Bicell vessel in units of 50 islets. Postwarm/thaw survival rates of the islets were 68.1% ± 5.9%, 64.1% ± 3.5%, and 47.7 ± 1.2% following Cryotop vitrification, nylon mesh vitrification, and Bicell freezing, respectively (p < 0.05). Glucose-stimulated insulin secretion in the two vitrification groups (stimulus index [SI] = 3.1-3.9) was superior to that in the freezing group (SI = 0.8). Additional experiments involved scaling-up the cryopreservation process using the nylon mesh device in units of 10, 50, or 100 islets. Increased numbers of islets per device had no adverse effects on cryosurvival (58.6%-68.5%) or insulin secretion potential (SI = 2.8-4.2). As the nylon mesh device does not require the handling of individual islets with glass pipettes, pre- and postvitrification islet treatment is less complicated. Therefore, nylon mesh can serve as a simple cryodevice for the vitrification of large quantities of rat pancreatic islets.
胰岛移植的实际需求使得大量胰岛必须以简单便捷的方式长期冷冻保存。我们采用以下方法冷冻保存大鼠胰岛（平均直径范围为101 - 150μm）：使用Cryotop装置或ø = 57μm的尼龙网装置以10个胰岛为单位进行玻璃化冷冻，或使用Bicell容器以50个胰岛为单位进行传统冷冻。在Cryotop玻璃化冷冻、尼龙网玻璃化冷冻和Bicell冷冻后，胰岛复温/解冻后的存活率分别为68.1%±5.9%、64.1%±3.5%和47.7±1.2%（p < 0.05）。两个玻璃化冷冻组的葡萄糖刺激胰岛素分泌（刺激指数[SI]=3.1 - 3.9）优于冷冻组（SI = 0.8）。额外的实验涉及使用尼龙网装置以10、50或100个胰岛为单位扩大冷冻保存过程。每个装置中胰岛数量的增加对冷冻存活率（58.6% - 68.5%）或胰岛素分泌潜力（SI = 2.8 - 4.2）没有不利影响。由于尼龙网装置不需要用玻璃吸管处理单个胰岛，玻璃化冷冻前后的胰岛处理不太复杂。因此，尼龙网可作为一种简单的冷冻装置用于大量大鼠胰岛的玻璃化冷冻。
