Isolation, purification and functional identification of Syrian golden hamster islets.
- Author:
Kan SUN
1
;
Jia SUN
;
Hong CHEN
;
Hua ZHANG
;
De-hong CAI
Author Information
- Publication Type:Journal Article
- MeSH: Animals; Cell Separation; methods; Cricetinae; Insulin; secretion; Islets of Langerhans; cytology; Male; Mesocricetus
- From: Journal of Southern Medical University 2009;29(9):1809-1811
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo establish the methods for isolation, purification and function identification of Syrian golden hamster islets.
METHODSThe Langerhans islets were isolated and purified from golden hamster pancreas by intra-ductal collagenase V perfusion and discontinuous Ficoll density gradient centrifugation. After isolation, the islet yield and purity were evaluated with DTZ staining. The islet function was assessed using glucose-stimulated insulin secretion test.
RESULTSThe total number of purified islets from one donor hamster was 359-/+35 islet equivalent (IEQ), with the purity and viability of the isolated islets of more than 90%. In response to glucose stimulations at 5.8 and 16.7 mmol/L, the secretion of insulin by the islets was 3.29-/+0.3 and 11.12-/+0.57 mU/L, respectively, showing a 2.28-fold higher insulin release by high-concentration than by low- concentration glucose stimulation (P<0.01).
CONCLUSIONThe methods of collagenase V digestion and gradient centrifugation result in high yield and high purity of the isolated hamster islets.