Utilization of rat and human sera to carry out incubation and perifusion of pancreatic islets.
Incubation of isolated pancreatic rat islets in a medium consisting of fresh rat or human sera and the measurement of insulin secretion after 60 min in the presence of 6.0-7.0 and 16.7 mM of glucose, were carried out. Perifusion experiments with isolated rat islets preincubated in the presence of rat or human sera and 0.2 mCi/mL of 45Ca2+ were also performed, and 45Ca2+ outflow rate and insulin secretion were analyzed. In both experiments, the usual islet responsiveness to glucose stimulation was preserved when compared with experiments in which Krebs-Henseleit (K-H) buffer was used as the medium. The results obtained demonstrate that both rat and human sera can be used as media for islets perifusion and incubation experiments, thus allowing for the effect(s) of circulating substances on insulin release to be examined.
el Razi
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Department of Physiology and Biophysics, University of São Paulo, Brazil.
Curi
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Carpinelli
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AR
D000328:Adult; D000818:Animals; D001769:Blood; D001774:Blood Chemical Analysis; D001786:Blood Glucose / Q000032:analysis; D002118:Calcium / Q000378:metabolism; D002132:Calcium Radioisotopes; D003470:Culture Media; D006801:Humans; D066298:In Vitro Techniques; D007328:Insulin / Q000378:metabolism; D000078790:Insulin Secretion; D007515:Islets of Langerhans / Q000502:physiology*; D008297:Male; D051381:Rats; D014021:Tissue Preservation / Q000379:methods*
D001786:Blood Glucose; D002132:Calcium Radioisotopes; D003470:Culture Media; D007328:Insulin; D002118:Calcium