Pancreatic Beta-Cell Purification by Altering FAD and NAD(P)H Metabolism

Figure 2

FAD fluorescence is altered by culture medium composition. Fluorescent patterns of dispersed islets preincubated in MEM, RPMI, CMRL, or DMEM-F12 medium (a). Analysis of glucagon-positive alpha cells present in the high fluorescent cell fraction following preincubation in MEM, RPMI, CMRL, or DMEM-F12 and subsequent sorting (b). Sorting of beta cells from the MEM incubated cell mixture resulted in a more than 95% pure beta-cell population, whereas in the nonbeta-cell fraction glucagon positive cells and a small amount of insulin positive cells were observed (c). Analysis of PP/somatostatin-positive cells present in the high and low fluorescent cell fractions following preincubation in MEM medium and subsequent sorting (d). Results are presented as the mean ± SEM, and statistical significance was determined using the Mann-Whitney 𝑈 test. 𝑃-values <.05 were considered to be statistically significant and are represented by (∗).