Isolated Islets Static Infusions or Perifusions

Isolation of murine pancreatic islets

Pancreatic islets are isolated from control or genetically-modified mice after digestion of the whole pancreas by ductal injection of collagenase. Islets are purified by hand-picking or by density centrifugation. Islet yield (50-200 islets/mouse pancreas) will strongly vary depending on mice age (optimal around 2-3 months), strain, and potential deleterious effects of the genetic manipulation on the pancreatic structure.

The number of mice should be discussed and will vary according to the experimental design and the mouse strain.

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Measurement of insulin secretion by pancreatic islets

Perifusions or static incubations of islets are performed to establish secretory responses to substrates such as glucose, to neurotransmitters such as acetylcholine, or to incretin factors such as GLP1 and GIP.

Perifusion of islets allows for dynamic characterization of glucose-stimulated insulin secretion. After the experiment, total islet insulin content is measured. Isolated islets can also be analyzed for second messengers such as cAMP.

The number of experiments should be discussed and will vary according to the experimental design and the mouse strain.

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References :

Preitner F, Ibberson M, Franklin I, Binnert C, Pende M, Gjinovci A, Hansotia T, Drucker DJ, Wollheim C, Burcelin R, Thorens B. Gluco-incretins control insulin secretion at multiple levels as revealed in mice lacking GLP-1 and GIP receptors. J Clin Invest. 2004 Feb;113(4):635-45.

Thorens B, Guillam MT, Beermann F, Burcelin R, Jaquet M. Transgenic reexpression of GLUT1 or GLUT2 in pancreatic beta cells rescues GLUT2-null mice from early death and restores normal glucose-stimulated insulin secretion. J Biol Chem. 2000 Aug 4;275(31):23751-8.

Guillam MT, Hummler E, Schaerer E, Yeh JI, Birnbaum MJ, Beermann F, Schmidt A, Deriaz N, Thorens B. Early diabetes and abnormal postnatal pancreatic islet development in mice lacking Glut-2. Nat Genet. 1997 Nov;17(3):327-30. Erratum in: Nat Genet 1997 Dec;17(4):503.