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重新审视小鼠模型中的胰岛分离:一种实用且有效的技术方案。

Revisiting pancreatic islet isolation in murine models: A practical and effective technical protocol.

机构信息

Laboratory of Morphometry, Metabolism, and Cardiovascular Diseases, Biomedical Center, Institute of Biology Roberto Alcantara Gomes, State University of Rio de Janeiro, Rio de Janeiro, Brazil.

Laboratory of Endocrine Physiology, Department of Physiological Sciences, Institute of Biology Roberto Alcantara Gomes, State University of Rio de Janeiro, Rio de Janeiro, Brazil.

出版信息

Physiol Rep. 2024 May;12(9):e16040. doi: 10.14814/phy2.16040.

Abstract

The endocrine pancreas is composed of clusters of cell groups called pancreatic islets. These cells are responsible for the synthesis and secretion of hormones crucial for glycemic homeostasis, such as insulin and glucagon. Therefore, these cells were the targets of many studies. One method to study and/or understand endocrine pancreatic physiology is the isolation of these islets and stimulation of hormone production using different concentrations of glucose, agonists, and/or antagonists of specific secretagogues and mimicking the stimulation of hormonal synthesis and secretion. Many researchers studied pancreatic physiology in murine models due to their ease of maintenance and rapid development. However, the isolation of pancreatic islets involves meticulous processes that may vary between rodent species. The present study describes a simple and effective technical protocol for isolating intact islets from mice and rats for use as a practical guide for researchers. The method involves digestion of the acinar parenchyma by intraductal collagenase. Isolated islets are suitable for in vitro endocrine secretion analyses, microscopy techniques, and biochemical analyses.

摘要

内分泌胰腺由称为胰岛的细胞群簇组成。这些细胞负责合成和分泌对血糖稳态至关重要的激素,如胰岛素和胰高血糖素。因此,这些细胞成为了许多研究的目标。研究内分泌胰腺生理学的一种方法是使用不同浓度的葡萄糖、激动剂和/或特定分泌刺激物的拮抗剂分离这些胰岛,并刺激激素产生,模拟激素合成和分泌的刺激。由于其易于维持和快速发育,许多研究人员在小鼠模型中研究胰腺生理学。然而,胰岛的分离涉及到精细的过程,不同的啮齿动物物种可能会有所不同。本研究描述了一种从小鼠和大鼠中分离完整胰岛的简单有效的技术方案,可作为研究人员的实用指南。该方法涉及通过胰管内胶原酶消化腺泡实质。分离的胰岛适合进行体外内分泌分泌分析、显微镜技术和生化分析。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e89a/11082087/cfde29202944/PHY2-12-e16040-g003.jpg

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