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A Practical Guide to Rodent Islet Isolation and Assessment Revisited

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...

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Autores principales: Corbin, Kathryn L., West, Hannah L., Brodsky, Samantha, Whitticar, Nicholas B., Koch, William J., Nunemaker, Craig S.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7919091/
https://www.ncbi.nlm.nih.gov/pubmed/33641671
http://dx.doi.org/10.1186/s12575-021-00143-x
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author Corbin, Kathryn L.
West, Hannah L.
Brodsky, Samantha
Whitticar, Nicholas B.
Koch, William J.
Nunemaker, Craig S.
author_facet Corbin, Kathryn L.
West, Hannah L.
Brodsky, Samantha
Whitticar, Nicholas B.
Koch, William J.
Nunemaker, Craig S.
author_sort Corbin, Kathryn L.
collection PubMed
description 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. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1186/s12575-021-00143-x.
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spelling pubmed-79190912021-03-02 A Practical Guide to Rodent Islet Isolation and Assessment Revisited Corbin, Kathryn L. West, Hannah L. Brodsky, Samantha Whitticar, Nicholas B. Koch, William J. Nunemaker, Craig S. Biol Proced Online Review 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. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1186/s12575-021-00143-x. BioMed Central 2021-03-01 /pmc/articles/PMC7919091/ /pubmed/33641671 http://dx.doi.org/10.1186/s12575-021-00143-x Text en © The Author(s) 2021 Open AccessThis article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated in a credit line to the data.
spellingShingle Review
Corbin, Kathryn L.
West, Hannah L.
Brodsky, Samantha
Whitticar, Nicholas B.
Koch, William J.
Nunemaker, Craig S.
A Practical Guide to Rodent Islet Isolation and Assessment Revisited
title A Practical Guide to Rodent Islet Isolation and Assessment Revisited
title_full A Practical Guide to Rodent Islet Isolation and Assessment Revisited
title_fullStr A Practical Guide to Rodent Islet Isolation and Assessment Revisited
title_full_unstemmed A Practical Guide to Rodent Islet Isolation and Assessment Revisited
title_short A Practical Guide to Rodent Islet Isolation and Assessment Revisited
title_sort practical guide to rodent islet isolation and assessment revisited
topic Review
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7919091/
https://www.ncbi.nlm.nih.gov/pubmed/33641671
http://dx.doi.org/10.1186/s12575-021-00143-x
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