4.4 Article

Observing Islet Function and Islet-Immune Cell Interactions in Live Pancreatic Tissue Slices

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JOURNAL OF VISUALIZED EXPERIMENTS
DOI: 10.3791/62207

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资金

  1. NIH [R01 DK123292, T32 DK108736, UC4 DK104194, UG3 DK122638, P01 AI042288]
  2. JDRF [nPOD: 5-SRA-2018-557-Q-R]
  3. Leona M. & Harry B. Helmsley Charitable Trust [2018PG-T1D053]

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Live pancreatic tissue slices offer a novel approach to studying islet physiology and function while preserving tissue viability, function, and underlying pathologies. This method allows for the maintenance of complex structures and cellular interactions for in-depth research on the pancreas.
Live pancreatic tissue slices allow for the study of islet physiology and function in the context of an intact islet microenvironment. Slices are prepared from live human and mouse pancreatic tissue embedded in agarose and cut using a vibratome. This method allows for the tissue to maintain viability and function in addition to preserving underlying pathologies such as type 1 (T1D) and type 2 diabetes (T2D). The slice method enables new directions in the study of the pancreas through the maintenance of the complex structures and various intercellular interactions that comprise the endocrine and exocrine tissues of the pancreas. This protocol demonstrates how to perform staining and time-lapse microscopy of live endogenous immune cells within pancreatic slices along with assessments of islet physiology. Further, this approach can be refined to discern immune cell populations specific for islet cell antigens using major histocompatibility complex-multimer reagents.

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