Department of Diabetes Immunology, Arthur Riggs Diabetes and Metabolism Research Institute;
Department of Diabetes Immunology, Arthur Riggs Diabetes and Metabolism Research Institute.
J Vis Exp. 2024 Mar 15(205). doi: 10.3791/66468.
It is crucial to study the human pancreas to understand the pathophysiological mechanisms associated with type 1 (T1D) and 2 diabetes (T2D) as well as the pancreas endocrine and exocrine physiology and interplay. Much has been learned from the study of isolated pancreatic islets, but this prevents examining their function and interactions in the context of the whole tissue. Pancreas slices provide a unique opportunity to explore the physiology of normal, inflamed, and structurally damaged islets within their native environment, in turn allowing the study of interactions between endocrine and exocrine compartments to better investigate the complex dynamics of pancreatic tissue. Thus, the adoption of the living pancreas slice platform represents a significant advancement in the field. This protocol describes how to generate living tissue slices from deceased organ donors by tissue embedding in agarose and vibratome slicing as well as their utilization to assess functional readouts such as dynamic secretion and live cell imaging.
研究人类胰腺对于理解与 1 型(T1D)和 2 型糖尿病(T2D)相关的病理生理机制以及胰腺内分泌和外分泌生理学及其相互作用至关重要。从对分离的胰岛的研究中已经了解到很多知识,但这阻止了在整个组织背景下检查它们的功能和相互作用。胰腺切片提供了一个独特的机会,可以在其天然环境中探索正常、炎症和结构受损胰岛的生理学,进而允许研究内分泌和外分泌隔室之间的相互作用,以更好地研究胰腺组织的复杂动力学。因此,活体胰腺切片平台的采用代表了该领域的重大进展。本协议描述了如何通过琼脂糖包埋和振动切片机切片从已故器官捐献者中生成活体组织切片,以及如何利用它们来评估动态分泌和活细胞成像等功能读数。