Corbin Kathryn L, West Hannah L, Brodsky Samantha, Whitticar Nicholas B, Koch William J, Nunemaker Craig S
Heritage College of Osteopathic Medicine, Ohio University, Athens, OH, USA.
Department of Biomedical Sciences, Ohio University, Athens, OH, USA.
Biol Proced Online. 2021 Mar 1;23(1):7. doi: 10.1186/s12575-021-00143-x.
Insufficient insulin secretion is a key component of both type 1 and type 2 diabetes. Since insulin is released by the islets of Langerhans, obtaining viable and functional islets is critical for research and transplantation. The effective and efficient isolation of these small islands of endocrine cells from the sea of exocrine tissue that is the rest of the pancreas is not necessarily simple or quick. Choosing and administering the digestive enzyme, separation of the islets from acinar tissue, and culture of islets are all things that must be considered. The purpose of this review is to provide a history of the development of islet isolation procedures and to serve as a practical guide to rodent islet research for newcomers to islet biology. We discuss key elements of mouse islet isolation including choosing collagenase, the digestion process, purification of islets using a density gradient, and islet culture conditions. In addition, this paper reviews techniques for assessing islet viability and function such as visual assessment, glucose-stimulated insulin secretion and intracellular calcium measurements. A detailed protocol is provided that describes a common method our laboratory uses to obtain viable and functional mouse islets for in vitro study. This review thus provides a strong foundation for successful procurement and purification of high-quality mouse islets for research purposes.
胰岛素分泌不足是1型和2型糖尿病的关键组成部分。由于胰岛素是由胰岛分泌的，因此获得有活力且功能正常的胰岛对于研究和移植至关重要。从胰腺其余部分的外分泌组织海洋中有效且高效地分离出这些小的内分泌细胞岛并非易事，也不一定很快。选择和使用消化酶、将胰岛与腺泡组织分离以及胰岛培养都是必须考虑的事项。本综述的目的是提供胰岛分离程序的发展历程，并为胰岛生物学新手进行啮齿动物胰岛研究提供实用指南。我们讨论了小鼠胰岛分离的关键要素，包括选择胶原酶、消化过程、使用密度梯度纯化胰岛以及胰岛培养条件。此外，本文还综述了评估胰岛活力和功能的技术，如视觉评估、葡萄糖刺激的胰岛素分泌和细胞内钙测量。提供了一份详细方案，描述了我们实验室用于获取有活力且功能正常的小鼠胰岛以进行体外研究的常用方法。因此，本综述为成功获取和纯化用于研究目的的高质量小鼠胰岛奠定了坚实基础。
