Formation of islet cell monolayers from fetal human and neonatal rat pancreatic islets: protein biosynthesis, insulin secretion, and binding of islet cell antibodies

Abstract

The phosphodiesterase inhibitor 3-isobutyl-1-methylxanthine (IBMX) has been used to promote monolayer formation in cultured islets isolated from fetal human or neonatal rat pancreas. Immunofluorescence with specific antiserum to insulin revealed B-cells in the outgrown monolayers. The rat islet cells were further characterized by their secretory and biosynthetic response to the action of IBMX. Both glucose-stimulated insulin release and recoverable insulin, i.e. intracellular insulin plus insulin secreted, were increased by the addition of IBMX (0.1 mmol/l) to the medium containing 10 mmol/l glucose. 3H-leucine incorporation into (pro)insulin was significantly higher following culture in 10 mmol/l glucose plus IBMX (0.1 and 1.0 mmol/l) than after cultivation with glucose alone. However, the percentage of (pro)insulin synthesized in relation to total protein synthesis was increased to a lesser extent after an acute incubation for 3 h at 1.5 mmol/l or in the absence of glucose. Moreover, the culture system employed in the present study proved to be useful for detecting islet cell antibodies bound to human as well as rat islet cells after exposure to islet cell antibody-positive sera.