A Practical Guide to Rodent Islet Isolation and Assessment

Biological Procedures Online

Jeffrey D Carter²  Stacey B Dula¹  Kathryn L Corbin²  Runpei Wu¹  Craig S Nunemaker² 

[1] Department of Medicine, Division of Endocrinology, University of Virginia, P.O. Box 801413, Charlottesville, VA, 22908, USA

[2] DERC Cell and Islet Isolation Core Facility, University of Virginia, Charlottesville, VA, USA

关键词: islet;    rat;    mouse;    rodent;    collagenase;    islet isolation;    isolation;    necrosis;    apoptosis;    insulin;    calcium;    transplantation;    cytokines;    beta cells;    beta-cell;    pancreatic islets;    islets of Langerhans;   

Others  :  797149
DOI  :  10.1007/s12575-009-9021-0

 received in 2009-06-18, accepted in 2009-10-22,  发布年份 2009

【 摘 要 】

Pancreatic islets of Langerhans secrete hormones that are vital to the regulation of blood glucose and are, therefore, a key focus of diabetes research. Purifying viable and functional islets from the pancreas for study is an intricate process. This review highlights the key elements involved with mouse and rat islet isolation, including choices of collagenase, the collagenase digestion process, purification of islets using a density gradient, and islet culture conditions. In addition, this paper reviews commonly used techniques for assessing islet viability and function, including visual assessment, fluorescent markers of cell death, glucose-stimulated insulin secretion, and intracellular calcium measurements. A detailed protocol is also included that describes a common method for rodent islet isolation that our laboratory uses to obtain viable and functional mouse islets for in vitro study of islet function, beta-cell physiology, and in vivo rodent islet transplantation. The purpose of this review is to serve as a resource and foundation for successfully procuring and purifying high-quality islets for research purposes.

【 授权许可】

   
2009 Carter et al.

【 图 表 】

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