A perifusion system for the study of insulin secretory dynamics of a clonal, Simian virus 40-transformed hamster pancreatic beta cell line (HIT cells) is described. After a change from glucose-free to higher glucose levels in the perifusate, insulin secretion increased rapidly in a dose-dependent manner. The pattern of glucose-stimulated insulin release was monophasic and was not sustained during a continued glucose stimulus. Perifusing the cells with low glucose (0.3 mg/ml) before a glucose stimulus of 3.5 mg/ml resulted in more rapid insulin release with lower peak secretory rates than those seen after a glucose-free period. The combined stimulus of high glucose and 100 μM 3-isobutyl-1-methylxanthine (IBMX), a phosphodiesterase inhibitor, significantly enhanced the acute insulin secretory response and also resulted in a biphasic secretory pattern that was sustained throughout the 60-min stimulation period. Insulin secretion stimulated by IBMX required a nonstimulatory level of glucose in the perifusing media, and, if this requirement was met, the immediate release of insulin was similar to that evoked by high glucose alone. High potassium (40 mM) also triggered a monophasic release of insulin. These studies demonstrate that glucose or high K+, which depolarizes the plasma membrane, and IBMX, an agent presumed to increase intracellular cyclic AMP levels, can signal the acute release of insulin from these beta cells. This cell line is a unique model system for studying the mechanism of insulin secretion.
Perifusion of a Clonal Cell Line of Simian Virus 40-Transformed Beta Cells: Insulin Secretory Dynamics in Response to Glucose, 3-Isobutyl-1-Methylxanthine, and Potassium
Ronald S Hill, A E Boyd; Perifusion of a Clonal Cell Line of Simian Virus 40-Transformed Beta Cells: Insulin Secretory Dynamics in Response to Glucose, 3-Isobutyl-1-Methylxanthine, and Potassium. Diabetes 1 February 1985; 34 (2): 115–120. https://doi.org/10.2337/diab.34.2.115
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