Department of Hepatopancreatobiliary Surgery, Shenzhen Institute of Translational Medicine, The First Affiliated Hospital of Shenzhen University, Shenzhen Second People's Hospital; Shenzhen Xenotransplantation Medical Engineering Research and Development Center, Shenzhen Institute of Translational Medicine, First Affiliated Hospital of Shenzhen University, Shenzhen Second People's Hospital.
Imaging Department, Shenzhen Institute of Translational Medicine, The First Affiliated Hospital of Shenzhen University, Shenzhen Second People's Hospital.
J Vis Exp. 2021 Oct 21(176). doi: 10.3791/63017.
Type 1 diabetes mellitus (T1DM) is caused by autoimmune destruction of pancreatic β cells, which results in little or no insulin production. Islet transplantation plays an important role in the treatment of T1DM, with the improved glycometabolic control, the reduced progression of complications, the reduction of hypoglycemic episodes when compared with traditional insulin therapy. The results of phase III clinical trial also demonstrated the safety and efficacy of islet allotransplantation in T1DM. However, the shortage of pancreas donors limits its widespread use. Animals as a source of islets such as the pig offer an alternative choice. Because the architecture of the pig pancreas is different from the islets of mice or humans, the pig islet isolation procedure is still challenging. Since the translation of alternative porcine islet sources (xenogeneic) to the clinical setting for treating T1DM through cellular transplantation is of great importance, a cost-effective, standardized, and reproducible protocol for isolating porcine islets is urgently needed. This manuscript describes a simplified and cost-effective method to isolate and purify adult porcine islets based on the previous protocols that have successfully transplanted porcine islets to non-human primates. This will be a beginners guide without the use of specialized equipment such as a COBE 2991 Cell Processor.
1 型糖尿病（T1DM）是由胰岛β细胞的自身免疫性破坏引起的，导致胰岛素产生很少或没有。胰岛移植在 T1DM 的治疗中起着重要作用，与传统的胰岛素治疗相比，它改善了糖代谢控制，减缓了并发症的进展，减少了低血糖发作的次数。III 期临床试验的结果也证明了胰岛同种异体移植在 T1DM 中的安全性和有效性。然而，胰腺供体的短缺限制了其广泛应用。动物作为胰岛的来源，如猪，提供了另一种选择。由于猪胰腺的结构与小鼠或人类的胰岛不同，因此猪胰岛的分离过程仍然具有挑战性。由于通过细胞移植将替代的猪胰岛源（异种）转化为治疗 T1DM 的临床应用非常重要，因此迫切需要一种经济高效、标准化和可重复的方法来分离和纯化猪胰岛。本文描述了一种简化且经济高效的方法，基于已成功将猪胰岛移植到非人类灵长类动物的先前方案来分离和纯化成年猪胰岛。这将是一个没有使用 COBE 2991 细胞处理器等专业设备的入门指南。
