Kumagai-Braesch Makiko, Yao Ming, Ågren Nils, Karadagi Ahmad, Ericzon Bo-Göran, Domogatskaya Anna
Division of Transplantation Surgery, Department of Clinical Science, Intervention and Technology, Karolinska Institutet, Stockholm, Sweden.
Department of Transplantation Surgery, Karolinska University Hospital, Huddinge, Sweden.
PLoS One. 2025 Jun 3;20(6):e0325421. doi: 10.1371/journal.pone.0325421. eCollection 2025.
To understand the mechanisms of diabetes and develop novel drugs, various animal diabetes models as well as in vitro experimental systems have been established. Using isolated islets in vitro is associated with several problems: (1) discontinued blood microcirculation causes ischemia and central necrosis in isolated islets, (2) dissociation of islets into single cells reduces the total number of cells and impairs endocrine function. Here, we aim to establish a novel experimental method using isolated islets to characterize cell physiology and drug effects. We have developed a method to flatten islets into monolayers by culturing small, whole, non-disrupted mouse islets on laminin-521. In this culture, islet cells remain intact, and cell-cell junctions are not broken mechanically. Small mouse islets were cultured in a 96-well plate coated with laminin-521 for six days and then exposed to various concentrations of streptozotocin (STZ) for 24 hours. Monolayer islets exposed to 0.6 mM STZ-induced a metabolic diabetes-like phenotype, i.e., islet cell death, predominantly of beta cells, causing a shift in alpha-to-beta cell ratio. The stimulation index in a glucose-stimulated insulin secretion assay was reduced in STZ-treated islets. However, insulin production per beta cell did not change significantly. The monolayer whole islet assay format allows the extraction of quantitative data regarding endocrine cell populations, "endocrine sensitivity" (stimulation index), and "endocrine power" (amount of hormone secreted per cell). It is a novel, versatile, user-friendly, semi-automated, and financially attractive experimental platform.
为了理解糖尿病的发病机制并开发新型药物，已经建立了各种动物糖尿病模型以及体外实验系统。在体外使用分离的胰岛存在几个问题：（1）中断的血液循环会导致分离的胰岛出现缺血和中央坏死，（2）胰岛解离成单个细胞会减少细胞总数并损害内分泌功能。在此，我们旨在建立一种使用分离的胰岛来表征细胞生理学和药物作用的新型实验方法。我们开发了一种方法，通过在层粘连蛋白-521上培养小的、完整的、未破坏的小鼠胰岛，将胰岛平铺成单层。在这种培养中，胰岛细胞保持完整，细胞间连接不会被机械破坏。将小的小鼠胰岛在涂有层粘连蛋白-521的96孔板中培养6天，然后暴露于不同浓度的链脲佐菌素（STZ）24小时。暴露于0.6 mM STZ的单层胰岛诱导出代谢性糖尿病样表型，即胰岛细胞死亡，主要是β细胞死亡，导致α细胞与β细胞比例发生变化。在葡萄糖刺激的胰岛素分泌试验中，STZ处理的胰岛的刺激指数降低。然而，每个β细胞的胰岛素产量没有显著变化。单层全胰岛检测形式允许提取关于内分泌细胞群体、“内分泌敏感性”（刺激指数）和“内分泌能力”（每个细胞分泌的激素量）的定量数据。它是一个新颖、通用、用户友好、半自动且经济上有吸引力的实验平台。
