A Practical Guide to Rodent Islet Isolation and Assessment Revisited

Biological Procedures Online

Nicholas B. Whitticar¹  Samantha Brodsky²  Kathryn L. Corbin²  Craig S. Nunemaker³  William J. Koch⁴  Hannah L. West⁵ 

[1]Department of Biomedical Sciences, Ohio University, Athens, OH, USA

[2]Translational Biomedical Sciences Program, Graduate College, Ohio University, Athens, OH, USA

[3]Heritage College of Osteopathic Medicine, Ohio University, Athens, OH, USA

[4]Department of Biomedical Sciences, Ohio University, Athens, OH, USA

[5]Heritage College of Osteopathic Medicine, Ohio University, Athens, OH, USA

[6]Department of Biomedical Sciences, Ohio University, Athens, OH, USA

[7]Diabetes Institute, Ohio University, Athens, OH, USA

[8]Heritage College of Osteopathic Medicine, Ohio University, Athens, OH, USA

[9]Department of Biomedical Sciences, Ohio University, Athens, OH, USA

[10]Translational Biomedical Sciences Program, Graduate College, Ohio University, Athens, OH, USA

[11]Honors Tutorial College, Ohio University, Athens, OH, USA

关键词: Islets of Langerhans;    Isolation;    Beta cell;    Collagenase;    Density gradient;    Neonatal;    Calcium;    Insulin secretion;   

DOI  :  10.1186/s12575-021-00143-x

来源: Springer

【 摘 要 】

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.

【 授权许可】

CC BY