Protocol for quantifying SA-β-gal activity as a measure of senescence in islets of a mouse model of type 1 diabetes
Summary: A subpopulation of pancreatic beta cells becomes senescent during type 1 diabetes (T1D) progression, and removal of these populations protects against T1D in mice. Here, we present a protocol to measure senescence in murine pancreatic islet cells through analysis of senescence-associated β-...
| Main Authors: | , , |
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| Format: | Article |
| Language: | English |
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Elsevier
2024-03-01
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| Series: | STAR Protocols |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S2666166724000881 |
| _version_ | 1827336619633410048 |
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| author | Hugo Lee Peter J. Thompson Feyza Engin |
| author_facet | Hugo Lee Peter J. Thompson Feyza Engin |
| author_sort | Hugo Lee |
| collection | DOAJ |
| description | Summary: A subpopulation of pancreatic beta cells becomes senescent during type 1 diabetes (T1D) progression, and removal of these populations protects against T1D in mice. Here, we present a protocol to measure senescence in murine pancreatic islet cells through analysis of senescence-associated β-galactosidase activity. We describe steps for staining with the fluorogenic substrate C12FDG and analysis by flow cytometry. Increased cell size is another marker of senescence and can also be concurrently measured in the same experiment.For complete details on the use and execution of this protocol, please refer to Lee et al.1 and Helman et al.2 : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics. |
| first_indexed | 2024-03-07T18:35:35Z |
| format | Article |
| id | doaj.art-bcf47eb9f10e430dafad249751dcd9f7 |
| institution | Directory Open Access Journal |
| issn | 2666-1667 |
| language | English |
| last_indexed | 2024-03-07T18:35:35Z |
| publishDate | 2024-03-01 |
| publisher | Elsevier |
| record_format | Article |
| series | STAR Protocols |
| spelling | doaj.art-bcf47eb9f10e430dafad249751dcd9f72024-03-02T04:54:58ZengElsevierSTAR Protocols2666-16672024-03-0151102923Protocol for quantifying SA-β-gal activity as a measure of senescence in islets of a mouse model of type 1 diabetesHugo Lee0Peter J. Thompson1Feyza Engin2Department of Biomolecular Chemistry, University of Wisconsin-Madison, School of Medicine and Public Health, Madison, WI 53706, USA; Corresponding authorDepartment of Physiology & Pathophysiology, Rady Faculty of Health Sciences, University of Manitoba, Winnipeg, MB RT3 2N2, CanadaDepartment of Biomolecular Chemistry, University of Wisconsin-Madison, School of Medicine and Public Health, Madison, WI 53706, USA; Department of Medicine, Division of Endocrinology, Diabetes & Metabolism, University of Wisconsin-Madison, School of Medicine and Public Health, Madison, WI 53705, USA; Department of Cell and Regenerative Biology, Wisconsin Institute for Discovery, University of Wisconsin-Madison, School of Medicine and Public Health, Madison, WI 53705, USA; Corresponding authorSummary: A subpopulation of pancreatic beta cells becomes senescent during type 1 diabetes (T1D) progression, and removal of these populations protects against T1D in mice. Here, we present a protocol to measure senescence in murine pancreatic islet cells through analysis of senescence-associated β-galactosidase activity. We describe steps for staining with the fluorogenic substrate C12FDG and analysis by flow cytometry. Increased cell size is another marker of senescence and can also be concurrently measured in the same experiment.For complete details on the use and execution of this protocol, please refer to Lee et al.1 and Helman et al.2 : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.http://www.sciencedirect.com/science/article/pii/S2666166724000881Cell BiologyCell cultureCell isolationMetabolismModel Organisms |
| spellingShingle | Hugo Lee Peter J. Thompson Feyza Engin Protocol for quantifying SA-β-gal activity as a measure of senescence in islets of a mouse model of type 1 diabetes STAR Protocols Cell Biology Cell culture Cell isolation Metabolism Model Organisms |
| title | Protocol for quantifying SA-β-gal activity as a measure of senescence in islets of a mouse model of type 1 diabetes |
| title_full | Protocol for quantifying SA-β-gal activity as a measure of senescence in islets of a mouse model of type 1 diabetes |
| title_fullStr | Protocol for quantifying SA-β-gal activity as a measure of senescence in islets of a mouse model of type 1 diabetes |
| title_full_unstemmed | Protocol for quantifying SA-β-gal activity as a measure of senescence in islets of a mouse model of type 1 diabetes |
| title_short | Protocol for quantifying SA-β-gal activity as a measure of senescence in islets of a mouse model of type 1 diabetes |
| title_sort | protocol for quantifying sa β gal activity as a measure of senescence in islets of a mouse model of type 1 diabetes |
| topic | Cell Biology Cell culture Cell isolation Metabolism Model Organisms |
| url | http://www.sciencedirect.com/science/article/pii/S2666166724000881 |
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