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

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

STAR protocols (2021-08-20)
Yun-Xia Zhu, Yun-Cai Zhou, Yan Zhang, Peng Sun, Xiao-Ai Chang, Xiao Han
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).

MATERIALIEN
Produktnummer
Marke
Produktbeschreibung

Sigma-Aldrich
Natriumchlorid, ACS reagent, ≥99.0%
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Ammoniumhydroxid -Lösung, ACS reagent, 28.0-30.0% NH3 basis
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Kaliumchlorid, ACS reagent, 99.0-100.5%
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Histopaque®-1077, sterile-filtered, density: 1.077 g/mL
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Magnesiumsulfat, anhydrous, ReagentPlus®, ≥99.5%
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Magnesiumchlorid Hexahydrat, ACS reagent, 99.0-102.0%
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Calciumchlorid, anhydrous, BioReagent, suitable for insect cell culture, suitable for plant cell culture, ≥96.0%
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Collagenase aus Clostridium histolyticum, Type V, ≥1 FALGPA units/mg solid, >125 CDU/mg solid
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α-D-Glucose, anhydrous, 96%
Sigma-Aldrich
Dithizon, Practical Grade