Pular para o conteúdo
Merck
  • 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
RESUMO

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).

MATERIAIS
Número do produto
Marca
Descrição do produto

Sigma-Aldrich
Cloreto de sódio, ACS reagent, ≥99.0%
Sigma-Aldrich
Hidróxido de amônio, ACS reagent, 28.0-30.0% NH3 basis
Sigma-Aldrich
Cloreto de potássio, ACS reagent, 99.0-100.5%
Sigma-Aldrich
Histopaque®-1077, sterile-filtered, density: 1.077 g/mL
Sigma-Aldrich
Sulfato de magnésio, anhydrous, ReagentPlus®, ≥99.5%
Sigma-Aldrich
Cloreto de magnésio, ACS reagent, 99.0-102.0%
Sigma-Aldrich
Calcium chloride, anhydrous, BioReagent, suitable for insect cell culture, suitable for plant cell culture, ≥96.0%
Sigma-Aldrich
Álcool etílico, puro, 200 proof, anhydrous, ≥99.5%
Sigma-Aldrich
Colagenase, Type V, ≥1 FALGPA units/mg solid, >125 CDU/mg solid
Sigma-Aldrich
α-D-Glucose, anhydrous, 96%
Sigma-Aldrich
Dithizone, Practical Grade