Summary
Isolated adult rat pancreatic islets were dispersed into single cells and cultured free-floating for 3 to 4 d, during which time islet cells reaggregated spontaneously into spherical clusters or pseudoislets. The gross morphology of these tissues resembled nondissociated islets. Electron microscopy revealed well-preserved cell ultrastructure and intercellular membrane connections. Immunofluorescent localization of islet cell types showed that A cells tended to be peripherally distributed around a B cell core, with D cells scattered throughtout the aggregate, mass. The dynamics of insulin release from pseudoislets were evaluated in vitro by perifusion techniques. Pseudoislets exhibited clear biphasic dose-dependent insulin responses to 30 min glucose stimulation over the range 5.5 to 30 mM. Repeated 2-min pulses with 22 mM glucose elicited brief monophasic spikes of insulin release of, consistent magnitude.l-Arginine (5 to 20 mM) evoked biphasic insulin release but these responses were not dose-dependent. These data indicate that islet cells reaggregate into structures with close morphologic similarities to intact islets, and that pseudoislet B cells continue to secrete insulin in response to nutrient secretagogues, comparable to that seen with islets in vitro and in situ.
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This work was supported by grants from the Medical Research Council of New Zealand. D. W. H. was the recipient of a Novo Diabetes Research Scholarship.
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Hopcroft, D.W., Mason, D.R. & Scott, R.S. Insulin secretion from perifused rat pancreatic pseudoislets. In Vitro Cell Dev Biol 21, 421–427 (1985). https://doi.org/10.1007/BF02620828
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DOI: https://doi.org/10.1007/BF02620828