Islets were isolated with collagenase P (0.65 mg/ml). For some [Ca2+]c experiments, isolated islets were further dissociated with trypsin–EDTA (0.25%) to obtain dispersed cells, which were then plated on coverslips. Islets and isolated cells were cultured for up to three days (one day for perifusion experiments and one to three days for [Ca2+]c measurements) in RPMI 1640 medium containing 7 mM glucose and 10% heat-inactivated FBS.

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