Isolation and Purification of Rodent Pancreatic Islets 2 of Langerhans

O'Dowd, Jacqueline F. and Stocker, Claire J. (2020) Isolation and Purification of Rodent Pancreatic Islets 2 of Langerhans. In: Type 2 Diabetes : Methods and Protocols. Methods in Molecular Biology (2076). Humana Press. ISBN 978-1-4939-9880-7 (In Press)

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Abstract

This chapter describes the detailed protocol for the isolation and purification of islets of Langerhans from 6 AU1 rodent pancreas using collagenase digestion. The first step of the process is to separate and isolate the 7 insulin-producing islets of Langerhans from the rest of the pancreas. The pancreas is excised from the 8 animal, trimmed of nonpancreatic tissues before being inflated and chopped into small pieces. The 9 connective tissue is then broken down with a collagenase enzyme solution to selectively digest the bulk 10 of the exocrine tissue while leaving the endocrine islets intact and separated from their surrounding 11 non-islet tissue. Once this process is completed, the islets of Langerhans are separated from the remaining 12 mixture by centrifugation and purified by the means of hand picking. Once isolated, the subsequent islets 13 can be used for several varied experimental processes, including transplantation, the study of pathophysio- 14 logical mechanisms in diabetic conditions, and in the screening of novel therapeutic approaches in phar- 15 macological research.

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