Protocol for in vivo and ex vivo assessments of glucose-stimulated insulin secretion in mouse islet β cells

Pancreatic islet β cells secrete insulin in a biphasic manner when sensing high blood glucose level. This protocol describes the evaluation of different phases of insulin secretion, as well as basal, glucose-stimulated and total insulin secretion abilities, thereby enabling precise assessment of β c...

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Veröffentlicht in:STAR protocols 2021-09, Vol.2 (3), p.100728-100728, Article 100728
Hauptverfasser: Zhu, Yun-Xia, Zhou, Yun-Cai, Zhang, Yan, Sun, Peng, Chang, Xiao-Ai, Han, Xiao
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Sprache:eng
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Zusammenfassung:Pancreatic islet β cells secrete insulin in a biphasic manner when sensing high blood glucose level. This protocol describes the evaluation of different phases of insulin secretion, as well as basal, glucose-stimulated and total insulin secretion abilities, thereby enabling precise assessment of β cell function both in vivo and ex vivo. The in vivo assay consists of intravenous tube imbedding surgery and hyperglycemic clamp. The ex vivo assay consists of islet isolation, dynamic perfusion and static immersion. For complete details on the use and execution of this protocol, please refer to Sun et al. (2021). [Display omitted] •Glucose-stimulated biphasic insulin secretion (GSIS) in mouse islet β cells•Optimized and detailed tube embedding surgery for in vivo GSIS assay•Careful islet isolation and verification is essential for ex vivo GSIS assay•Adaptable procedure to monitor both human and murine islet β-cell function ex vivo Pancreatic islet β cells secrete insulin in a biphasic manner when sensing high blood glucose level. This protocol describes the evaluation of different phases of insulin secretion, as well as basal, glucose-stimulated and total insulin secretion abilities, thereby enabling precise assessment of β cell function both in vivo and ex vivo. The in vivo assay consists of intravenous tube imbedding surgery and hyperglycemic clamp. The ex vivo assay consists of islet isolation, dynamic perfusion and static immersion.
ISSN:2666-1667
2666-1667
DOI:10.1016/j.xpro.2021.100728