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，以使糖尿病大鼠模型中的血糖水平正常化。根据NM设备上的最小体积冷却规程或SF海绵盘上的固体表面玻璃化规程，将一百只大鼠胰岛冷冻保存。加热后胰岛的葡萄糖刺激的胰岛素分泌（GSIS）分析中的回收率（97.1％对93.8％），生存力（77.9％对74.4％）和刺激指数（4.7对4.2）在NM玻璃化组和SF玻璃化组之间具有可比性。新鲜对照胰岛的活力和刺激指数分别确定为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的肾囊下方（血糖水平> 350 mg / dl）。移植后3周内，在受体大鼠中观察到正常血糖（<200 mg / dl）的获得（80.0–83.3％）。在比较组中，第30天和第60天的腹膜内葡萄糖耐量测试显示对治愈大鼠的葡萄糖摄取具有相似的2小时反应。此外，第70天肾切除术通过随后的糖尿病逆转和组织学评估证实了移植物的成功植入。因此，事实证明，在NM设备或SF海绵盘上玻璃化加热的大量大鼠胰岛在体内和体外均具有完全的功能，