A buffer temperature controlled perifusion system to study temperature dependence and kinetics of insulin secretion in MIN6 pseudoislets

Journal of Pharmacological and Toxicological Methods
Martin B Brenner, Hans-Juergen Mest

Abstract

The perifusion of pancreatic islets is a well-known method to investigate the kinetics of insulin secretion. Nevertheless, little interest has been attributed to a precise temperature control in perifusion systems. Insulin secretion from MIN6 cells, cultured as monolayers, differs substantially from pancreatic islets, at least partly due to missing beta-to-beta cell contacts. These cellular contacts are abundant in MIN6 pseudoislets, which show a more pronounced glucose-induced insulin release. Here, a perifusion system that directly and dynamically controls the perifusion buffer temperature inside the reaction chamber is described. Additionally, the influence of small temperature changes, glucagon-like peptide 1 (GLP-1) and tolbutamide on insulin release from MIN6 pseudoislets is examined. MIN6 cells were cultured in suspension culture dishes to generate MIN6 pseudoislets. The pseudoislets were perifused using a newly developed 12-channel perifusion system. The buffer temperature inside the reaction chambers was dynamically controlled by a programmable proportional plus integral plus differential (PID) controller. Insulin was determined by radioimmunoassay. After adjusting the PID controller, the temperature inside the reactio...Continue Reading

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Citations

Jul 20, 2010·Biochemical and Biophysical Research Communications·Catriona KellyNeville H McClenaghan
Dec 20, 2007·Cell Transplantation·Kristen E La FlammeTejal A Desai
Jan 20, 2018·American Journal of Physiology. Endocrinology and Metabolism·Michael G SpeliosEitan M Akirav
Oct 26, 2005·Diabetes Technology & Therapeutics·Kristen E La FlammeTejal A Desai
Sep 7, 2013·Journal of Proteome Research·Azazul ChowdhuryPeter Bergsten

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