1] Deutsche Forschungsgemeinschaft (DFG) Research Center for Regenerative Therapies Dresden (CRTD), Technische Universität Dresden, Dresden, Germany. [2] Paul Langerhans Institute Dresden, University Hospital Carl Gustav Carus, Technische Universität Dresden, Dresden, Germany. [3] German Center for Diabetes Research (DZD), Dresden, Germany.
Max Planck Institute of Neurobiology, Martinsried, Germany.
Nat Protoc. 2014 Dec;9(12):2809-22. doi: 10.1038/nprot.2014.195. Epub 2014 Nov 13.
Studies on the cellular function of the pancreas are typically performed in vitro on its isolated functional units, the endocrine islets of Langerhans and the exocrine acini. However, these approaches are hampered by preparation-induced changes of cell physiology and the lack of an intact surrounding. We present here a detailed protocol for the preparation of pancreas tissue slices. This procedure is less damaging to the tissue and faster than alternative approaches, and it enables the in situ study of pancreatic endocrine and exocrine cell physiology in a conserved environment. Pancreas tissue slices facilitate the investigation of cellular mechanisms underlying the function, pathology and interaction of the endocrine and exocrine components of the pancreas. We provide examples for several experimental applications of pancreas tissue slices to study various aspects of pancreas cell biology. Furthermore, we describe the preparation of human and porcine pancreas tissue slices for the validation and translation of research findings obtained in the mouse model. Preparation of pancreas tissue slices according to the protocol described here takes less than 45 min from tissue preparation to receipt of the first slices.
研究胰腺的细胞功能通常在体外进行,使用其分离的功能单位,即胰岛和外分泌腺泡。然而,这些方法受到制备诱导的细胞生理学变化和缺乏完整环境的限制。我们在此介绍了一种详细的胰腺组织切片制备方案。该方法对组织的损伤较小,比其他方法更快,并且能够在保存环境中原位研究胰腺内分泌和外分泌细胞的生理学。胰腺组织切片有助于研究胰腺内分泌和外分泌成分的功能、病理学和相互作用的细胞机制。我们提供了几个胰腺组织切片的实验应用实例,以研究胰腺细胞生物学的各个方面。此外,我们还描述了人胰腺和猪胰腺组织切片的制备方法,用于验证和转化从小鼠模型中获得的研究结果。根据这里描述的方案制备胰腺组织切片,从组织准备到收到第一片切片不到 45 分钟。