Autofluorescence-activated cell sorting of pancreatic islet cells: Purification of insulin-containing B-cells according to glucose-induced changes in cellular redox state

Abstract

Autofluorescence-activated cell sorting can be employed for the subfractionation of insulin-containing islet B-cells according to their responsiveness to their physiologic stimulus, glucose. The method utilizes a flow cytometric detection of the rapid variations in endogenous NAD (P) H - and FAD - fluorescence after exposure to 20 mM glucose. Under these conditions, a two-fold increase in NAD (P) H and a 40% decrease in FAD was observed in more than 75% of B-cells isolated from fed normal rats. The technique makes it possible to study the metabolic behaviour of the B-cell population in (physio)pathological conditions of impaired glucose-induced insulin release; the availability of functionally homogenous B-cell preparations facilitates studies on stimulus-secretion coupling. In view of the universal role of the cellular metabolic redox state in cell regulation, it is suggested that similar techniques can be developed for the metabolic analysis of other cell types and for their purification according to their responsiveness to specific stimuli.