Technical Reference #1853

1853.

Glucose Suppression of Glucagon Secretion METABOLIC AND CALCIUM RESPONSES FROM alpha-CELLS IN INTACT MOUSE PANCREATIC ISLETS Sylvain J. Le Marchand and David W. Piston, Vanderbilt University, The Journal of Biological Chemistry, 285(1853), (2010)
Link To Paper

Materials & Methods:
Islet Isolation and Culture—Male mice (2–6-month-old) were anesthetized by intraperitoneal injection of 0.05 ml ketamine/xylazine (VedCo St. Joseph MO) at 80 and 20 mg/ml respectively. We used a modified version of the collagenase digestion method described elsewhere (36). Islets were maintained in islet medium composed of RPMI 1640 medium supplemented with 10% fetal bovine serum penicillin (100 units/ml) streptomycin (100 g/ml) and glucose (11 mM) at 37 °C under 5% humidifiedCO2. For imaging purposes isolated islets were attached on gelatin (0.1%)-coated 35-mm glass-bottomed dishes (MatTek Corp. Ashland MA).

Microscopic Technique
Immunofluorescence

Cell Type(s)
alpha-cells - pancreatic islets