Paul Langerhans Institute Dresden (PLID) of the Helmholtz Zentrum München at the University Clinic Carl Gustav Carus of Technische Universität Dresden, Helmholtz Zentrum München, Neuherberg, Germany.
Institute of Physiology, Faculty of Medicine, Technische Universität Dresden, Dresden, Germany.
Methods Mol Biol. 2020;2128:301-312. doi: 10.1007/978-1-0716-0385-7_20.
Studies on islet of Langerhans physiology are crucial to understand the role of the endocrine pancreas in diabetes pathogenesis and the development of new therapeutic approaches. However, so far most research addressing islet of Langerhans biology relies on islets obtained via enzymatic isolation from the pancreas, which is known to cause mechanical and chemical stress, thus having a major impact on islet cell physiology. To circumvent the limitations of islet isolation, we have pioneered a platform for the study of islet physiology using the pancreas tissue slice technique. This approach allows to explore the detailed three-dimensional morphology of intact pancreatic tissue at a cellular level and to investigate islet cell function under near-physiological conditions. The described procedure is less damaging and faster than alternative approaches and particularly advantageous for studying infiltrated and structurally damaged islets. Furthermore, pancreas tissue slices have proven valuable for acute studies of endocrine as well as exocrine cell physiology in their conserved natural environment. We here provide a detailed protocol for the preparation of mouse pancreas tissue slices, the assessment of slice viability, and the study of pancreas cell physiology by hormone secretion and immunofluorescence staining.
关于胰岛生理学的研究对于理解内分泌胰腺在糖尿病发病机制和新治疗方法发展中的作用至关重要。然而,到目前为止,大多数涉及胰岛生物学的研究都依赖于从胰腺中通过酶分离获得的胰岛,这已知会引起机械和化学应激,从而对胰岛细胞的生理学产生重大影响。为了规避胰岛分离的局限性,我们开创了一种使用胰腺组织切片技术研究胰岛生理学的平台。这种方法可以在细胞水平上探索完整胰腺组织的详细三维形态,并在接近生理的条件下研究胰岛细胞功能。与替代方法相比,所描述的程序对组织的损伤更小、速度更快,特别有利于研究浸润和结构受损的胰岛。此外,胰腺组织切片已被证明对于在其保守的自然环境中研究内分泌和外分泌细胞生理学非常有价值。在这里,我们提供了详细的 protocol,用于制备小鼠胰腺组织切片,评估切片活力,并通过激素分泌和免疫荧光染色研究胰腺细胞生理学。