ABSTRACT

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.

摘要

我们报告了在尼龙网 (NM) 装置或丝素蛋白 (SF) 海绵盘上玻璃化加热的大鼠胰岛的适应性，用于使糖尿病大鼠模型的血糖水平正常化。根据 NM 设备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冷冻保存一百只大鼠胰岛。加热后胰岛的葡萄糖刺激胰岛素分泌 (GSIS) 试验的回收率 (97.1% vs. 93.8%)、存活率 (77.9% vs. 74.4%) 和刺激指数 (4.7 vs. 4.2) 分别为NM 玻璃化和 SF 玻璃化组之间的可比性。新鲜对照胰岛的存活率和刺激指数分别为 97.5% 和 6.5。将来自 NM 或 SF 玻璃化组或新鲜对照组的 800 个胰岛移植到链脲佐菌素诱导的糖尿病大鼠 (血糖水平 > 350 mg/dl) 的肾囊下。移植后 3 周内，在受体大鼠 (80.0–83.3%) 中观察到血糖正常 (< 200 mg/dl)。在第 30 天和第 60 天进行的腹腔葡萄糖耐量试验显示，比较组中治愈的大鼠对葡萄糖摄取的 2 小时反应相似。此外，通过随后的糖尿病逆转和组织学评估，第 70 天肾切除术证实了移植物的成功植入。因此，在 NM 装置或 SF 海绵盘上玻璃化加热的大量大鼠胰岛被证明在体外和体内都具有充分的功能，