O'Dowd Jacqueline F
Clore Laboratory, University of Buckingham, Hunter Street, Buckingham, Buckinghamshire, MK18 1EG, UK.
Methods Mol Biol. 2009;560:37-42. doi: 10.1007/978-1-59745-448-3_3.
This chapter describes the detailed protocol for the isolation and purification of islets of Langerhans from rodent pancreas using collagenase digestion. The first step of the process is to separate and isolate the insulin-producing islets of Langerhans from the rest of the pancreas. The pancreas is excised from the animal, trimmed of non-pancreatic tissues before being inflated and chopped into small pieces. The connective tissue is then broken down with a collagenase enzyme solution to selectively digest the bulk of the exocrine tissue while leaving the endocrine islets intact and separated from their surrounding non-islet tissue. Once this process is completed, the islets of Langerhans are separated from the remaining mixture by centrifugation and purified by the means of hand picking. Once isolated, the subsequent islets can be used for a number of experimental processes.
本章介绍了使用胶原酶消化法从啮齿动物胰腺中分离和纯化胰岛的详细方案。该过程的第一步是将产生胰岛素的胰岛从胰腺的其余部分分离出来。从动物体内取出胰腺,去除非胰腺组织,然后进行充气并切成小块。接着用胶原酶溶液分解结缔组织,以选择性地消化大部分外分泌组织,同时使内分泌胰岛保持完整,并与周围的非胰岛组织分离。一旦这个过程完成,通过离心将胰岛从剩余混合物中分离出来,并通过手工挑选进行纯化。一旦分离出来,随后的胰岛可用于许多实验过程。