Laboratory of Tissue Engineering and Organ Regeneration, Department of Surgery, University of Geneva, Geneva, Switzerland.
Cell Isolation and Transplantation Center, Department of Surgery, Geneva University Hospitals and University of Geneva, Geneva, Switzerland.
Methods Mol Biol. 2023;2592:37-60. doi: 10.1007/978-1-0716-2807-2_3.
Clinical islet transplantation (CIT) is an established noninvasive treatment for type I diabetes (T1D) and has demonstrated improved glycemic control, preventing the occurrence of severe hypoglycemia. However, CIT has several limitations, such as the need for multiple donors, lifelong immunosuppression, and suboptimal long-term graft function. Most of the transplanted islets are lost due to inflammation, ischemic damage, and delayed revascularization.Generation of organoids have gained increasing interest in regenerative medicine in recent years. In the context of beta-cell replacement, it offers a possibility to address limitations of CIT by allowing to produce uniform organoids from single or multiple cell types facilitating revascularization and anti-inflammatory and/or immunomodulatory protection. We have previously generated multicellular insulin-secreting organoids composed of islet cells and the human amniotic epithelial cells (hAECs). These 3D insulin-secreting structures demonstrated improved viability and function both in vitro and in vivo. Here we detail a stepwise methodology to generate insulin-secreting organoids using two different methods. In addition, quality assessment in vitro tests are also described.
临床胰岛移植（CIT）是一种成熟的治疗 1 型糖尿病（T1D）的非侵入性方法，已证明其可改善血糖控制，预防严重低血糖的发生。然而，CIT 存在几个局限性，例如需要多个供体、终身免疫抑制和移植胰岛的长期功能不理想。大多数移植的胰岛由于炎症、缺血性损伤和延迟再血管化而丢失。近年来，类器官的产生在再生医学中引起了越来越多的关注。在β细胞替代的背景下，它提供了一种通过允许从单个或多个细胞类型产生均匀的类器官来解决 CIT 局限性的可能性，从而促进再血管化和抗炎和/或免疫调节保护。我们之前已经生成了由胰岛细胞和人羊膜上皮细胞（hAEC）组成的多细胞胰岛素分泌类器官。这些 3D 胰岛素分泌结构在体外和体内都显示出了更好的活力和功能。在这里，我们详细描述了使用两种不同方法生成胰岛素分泌类器官的逐步方法。此外，还描述了体外质量评估测试。
