Graduate School of Science and Technology, Shinshu University , Ueda, Japan.
Center for Genetic Analysis of Behavior, National Institute for Physiological Sciences , Okazaki, Japan.
Islets. 2020 Nov 1;12(6):145-155. doi: 10.1080/19382014.2020.1849928. Epub 2020 Dec 8.
We report the adaptability of rat islets vitrified-warmed on nylon mesh (NM) device or silk fibroin (SF) sponge disc for the normalization of the blood glucose level in rat models of diabetes. One-hundred rat islets were cryopreserved according to a minimum volume cooling protocol on an NM device or a solid surface vitrification protocol on an SF sponge disc. The recovery rate (97.1% vs. 93.8%), the viability (77.9% vs. 74.4%), and the stimulation index (4.7 vs. 4.2) in glucose-stimulated insulin secretion (GSIS) assay of the post-warm islets were comparable between the NM vitrification and the SF vitrification groups. The viability and the stimulation index of the fresh control islets were identified to be 97.5% and 6.5, respectively. Eight hundred islets from the NM or the SF vitrification group or the fresh control group were transplanted beneath the kidney capsule of a streptozotocin-induced diabetic rat (blood glucose level > 350 mg/dl). Within 3 weeks after transplantation, the acquisition of euglycemia (< 200 mg/dl) was observed in recipient rats (80.0-83.3%). An intraperitoneal glucose tolerance test on Day-30 and Day-60 showed similar 2-h responses to the glucose uptake of cured rats among the compared groups. Moreover, the successful engraftment of transplants was confirmed by the Day-70 nephrectomy through the subsequent diabetes reversal and histological evaluation. Thus, large quantities of rat islets vitrified-warmed on an NM device or an SF sponge disc were proven to be fully functional both in vitro and in vivo, due to the GSIS and syngeneic transplantation, respectively.
我们报告了在尼龙网（NM）装置或丝素蛋白（SF）海绵盘上对大鼠胰岛进行玻璃化-温热处理的适应性，以实现糖尿病大鼠模型的血糖水平正常化。将 100 个大鼠胰岛按照 NM 装置上的最小体积冷却方案或 SF 海绵盘上的固体表面玻璃化方案进行冷冻保存。在葡萄糖刺激胰岛素分泌（GSIS）测定中，NM 玻璃化和 SF 玻璃化组的温热后胰岛的回收率（97.1%对 93.8%）、活力（77.9%对 74.4%）和刺激指数（4.7 对 4.2）相似。新鲜对照胰岛的活力和刺激指数分别确定为 97.5%和 6.5。800 个胰岛分别来自 NM 或 SF 玻璃化组或新鲜对照组，移植到链脲佐菌素诱导的糖尿病大鼠（血糖水平>350mg/dl）的肾包膜下。移植后 3 周内，受体大鼠获得了正常血糖（<200mg/dl）（80.0-83.3%）。在第 30 天和第 60 天进行的腹腔内葡萄糖耐量试验显示，在接受治疗的大鼠中，各组的 2 小时葡萄糖摄取反应相似。此外，通过第 70 天的肾切除术，确认了移植的成功植入，随后出现糖尿病逆转和组织学评估。因此，大量在 NM 装置或 SF 海绵盘上玻璃化-温热的大鼠胰岛在体外和体内均被证明具有完全的功能，分别归因于 GSIS 和同种异体移植。
