关键词: Immunofluorescence staining In situ islet cell function Insulin secretion Live/dead staining Tissue slices

Mesh : Animals Cell Survival / physiology Fluorescent Antibody Technique / methods Histocytological Preparation Techniques / methods Insulin Secretion / physiology Islets of Langerhans / physiology Mice Microfluidic Analytical Techniques / methods Pancreas / cytology Tissue Culture Techniques / methods Tissue and Organ Harvesting

来  源:   DOI:10.1007/978-1-0716-0385-7_20

Abstract:
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.
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