Figure 2. Protocol for induction of glucotoxicity and assessment of responsiveness to physiologic glucosein pancreatic β-cells. Single β-cells isolated from rats were cultured under control and test conditions for 3 days. Subsequently, single β-cells were first incubated for 30 min in HKRB containing 2.8 mM glucose for stabilization and then subjected to [Ca2+]i measurements for functional assay.