First Department of Surgery, University of Fukui, Fukui 910-1193, Japan.
Department of Applied Chemistry and Biotechnology, Graduate School of Engineering, University of Fukui, Fukui 910-8507, Japan.
Nutrients. 2021 Aug 25;13(9):2959. doi: 10.3390/nu13092959.
Cryopreservation of pancreatic islets enables their long-term storage and subsequent transplantation; however, post-cryopreservation, islets viability, and functions are reduced to a significant extent. Islet is composed of five cells (α cell, β cell, δ cell, ε cell, and PP cell), and blood vessels that carry the nutrition. Freezing technology of the organization has not developed a good method. This paper is studied using a fructan which has been found to effectively freeze protect a material of the cell. Islet transplantation has been established as an effective means of treating patients with type 1 diabetes. In this study, we demonstrated the effectiveness of using a fructan on the cryopreserved islets by showing valid results for diabetes. Isolated rat islets were cryopreserved using phosphate-buffered saline (PBS) supplemented with different concentrations of fructan and/or dimethyl sulfoxide (DMSO) in FBS. The survival rates of the islets were estimated at different time intervals, and insulin secretion function was tested in vitro. Furthermore, the in vivo function was tested by syngeneic transplantation into streptozotocin-induced diabetic rats, and the grafts were analyzed histologically and immunohistochemically. Fructan significantly increased islet survival; 30% fructan led to survival rates of more than 90% on day 3, which was significantly higher than those of the DMSO groups ( < 0.05). For both fructan and DMSO, the survival showed dose dependence, with the highest rates observed for 30% fructan and 10% DMSO, respectively ( < 0.05). The fructan groups showed a significantly increased insulin secretion volume in comparison to the DMSO groups ( < 0.05). Furthermore, cell clusters of pancreatic islets were well maintained in the fructan group, whereas margin collapse and vacuolation were observed in the DMSO group. Three days after transplantation of pancreatic islets preserved with 30% fructan, the blood glucose levels of diabetic rats were restored to the normal range, and removal of transplanted pancreatic islets from the kidney led to a profound increase in blood glucose levels. Together, these results show that a fructan is effective at cryopreserving rat pancreatic islets for subsequent transplantation.
胰岛的低温保存使其能够长期储存和随后的移植;然而,在低温保存后,胰岛的活力和功能大大降低。胰岛由五种细胞(α细胞、β细胞、δ细胞、ε细胞和 PP 细胞)和携带营养的血管组成。组织的冷冻技术尚未开发出一种很好的方法。本文研究了一种已被发现可有效冷冻保护细胞的物质——果聚糖。胰岛移植已被确立为治疗 1 型糖尿病患者的有效手段。在这项研究中,我们通过展示对糖尿病有效的结果,证明了在低温保存的胰岛中使用果聚糖的有效性。使用磷酸盐缓冲盐水(PBS)培养分离的大鼠胰岛,其中添加了不同浓度的果聚糖和/或二甲亚砜(DMSO)在 FBS 中。在不同时间间隔估计胰岛的存活率,并在体外测试胰岛素分泌功能。此外,通过将胰岛移植到链脲佐菌素诱导的糖尿病大鼠体内进行体内功能测试,并对移植物进行组织学和免疫组织化学分析。果聚糖显著提高了胰岛的存活率;30%果聚糖在第 3 天导致存活率超过 90%,明显高于 DMSO 组(<0.05)。对于果聚糖和 DMSO,存活率均表现出剂量依赖性,分别观察到 30%果聚糖和 10%DMSO 的最高存活率(<0.05)。与 DMSO 组相比,果聚糖组的胰岛素分泌量显著增加(<0.05)。此外,在果聚糖组中,胰岛细胞簇得到了很好的维持,而在 DMSO 组中则观察到边缘塌陷和空泡化。在移植用 30%果聚糖保存的胰岛 3 天后,糖尿病大鼠的血糖水平恢复到正常范围,从肾脏中取出移植的胰岛会导致血糖水平显著升高。总之,这些结果表明,果聚糖在低温保存大鼠胰岛以进行随后的移植方面是有效的。