[Islet perifusion: a new method to evaluate first-phase insulin secretory function of beta-cell in vitro]. 2010

Jia-qing Shao, and Ping Gu, and Hong DU, and Bin Lu, and Li Peng, and Jie Li, and Yan-yan Wang, and Ming Zhao, and Jian Wang
Department of Endocrinology and Metabolism, Nanjing General Hospital of Nanjing Military Command, Nanjing 210002, China.

OBJECTIVE To set up islet perifusion system, a new method to evaluate first-phase insulin secretory function of beta-cell in vitro. METHODS Islet perifusion system was set up, including perifusion framework, waterbath system, infusion pump system, afferent system, islet capsule, and efferent system. Kinetics of insulin release in vitro was studied using the perifusion system. Pancreatic islets were isolated as mentioned above and used freshly after isolation. Size-matched 50 islets were placed in each column. Then the columns were gently closed with the top adaptors, immersed in vertical position and controlled temperature in the water bath at 37 degrees C. The perifusion medium was maintained at 37 degrees C in a water bath. And all columns were perifused in parallel at a flow rate of 0.5 ml/min with KRBB (2.8 mmol/L glucose) at 37 degrees C. After 60 min static incubation with KRBB (2.8 mmol/L glucose), the islets were stimulated in the continuous presence of a high concentration of 16.7 mmol/L glucose. Samples were collected every 20-second until 2 min, every 1 min until 5 min, thereafter every 5 min until 30 min. Samples were immediately stocked at -80 degrees C until further analysis. Insulin concentration was measured with an insulin RIA kit. RESULTS An Islet perifusion system was established successfully, and kinetic curves of insulin secretion of db/m and db/db mice were traced out. CONCLUSIONS An established islet perifusion system, could be widely used to evaluate insulin secretory function of beta-cell in in vitro research of diabetes.

UI MeSH Term Description Entries
D007328 Insulin A 51-amino acid pancreatic hormone that plays a major role in the regulation of glucose metabolism, directly by suppressing endogenous glucose production (GLYCOGENOLYSIS; GLUCONEOGENESIS) and indirectly by suppressing GLUCAGON secretion and LIPOLYSIS. Native insulin is a globular protein comprised of a zinc-coordinated hexamer. Each insulin monomer containing two chains, A (21 residues) and B (30 residues), linked by two disulfide bonds. Insulin is used as a drug to control insulin-dependent diabetes mellitus (DIABETES MELLITUS, TYPE 1). Iletin,Insulin A Chain,Insulin B Chain,Insulin, Regular,Novolin,Sodium Insulin,Soluble Insulin,Chain, Insulin B,Insulin, Sodium,Insulin, Soluble,Regular Insulin
D008297 Male Males
D008810 Mice, Inbred C57BL One of the first INBRED MOUSE STRAINS to be sequenced. This strain is commonly used as genetic background for transgenic mouse models. Refractory to many tumors, this strain is also preferred model for studying role of genetic variations in development of diseases. Mice, C57BL,Mouse, C57BL,Mouse, Inbred C57BL,C57BL Mice,C57BL Mice, Inbred,C57BL Mouse,C57BL Mouse, Inbred,Inbred C57BL Mice,Inbred C57BL Mouse
D010477 Perfusion Treatment process involving the injection of fluid into an organ or tissue. Perfusions
D000818 Animals Unicellular or multicellular, heterotrophic organisms, that have sensation and the power of voluntary movement. Under the older five kingdom paradigm, Animalia was one of the kingdoms. Under the modern three domain model, Animalia represents one of the many groups in the domain EUKARYOTA. Animal,Metazoa,Animalia
D046509 Tissue Culture Techniques A technique for maintaining or growing TISSUE in vitro, usually by DIFFUSION, perifusion, or PERFUSION. The tissue is cultured directly after removal from the host without being dispersed for cell culture. Culture Technique, Tissue,Culture Techniques, Tissue,Tissue Culture Technique
D050417 Insulin-Secreting Cells A type of pancreatic cell representing about 50-80% of the islet cells. Beta cells secrete INSULIN. Pancreatic beta Cells,beta Cells, Pancreatic,Pancreatic B Cells,B Cell, Pancreatic,B Cells, Pancreatic,Cell, Insulin-Secreting,Cells, Insulin-Secreting,Insulin Secreting Cells,Insulin-Secreting Cell,Pancreatic B Cell,Pancreatic beta Cell,beta Cell, Pancreatic
D051379 Mice The common name for the genus Mus. Mice, House,Mus,Mus musculus,Mice, Laboratory,Mouse,Mouse, House,Mouse, Laboratory,Mouse, Swiss,Mus domesticus,Mus musculus domesticus,Swiss Mice,House Mice,House Mouse,Laboratory Mice,Laboratory Mouse,Mice, Swiss,Swiss Mouse,domesticus, Mus musculus

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