Isolation and applications of prostate side population cells based on dye cycle violet efflux

This unit describes methods for digestion of human prostate clinical specimens and dye cycle violet (DCV) staining for identification, isolation, and quantitation of radiolabeled dihydrotestosterone (DHT) retention of side population cells. The principle of the side population assay is based on differential efflux of DCV, a cell-membrane-permeable fluorescent dye, by cells with high ATP-binding cassette (ABC) transporter activity. Cells with high ABC transporter activity that efflux DCV and fall in the lower left quadrant of a flow cytograph are designated as "side population" cells. This unit emphasizes tissue digestion, DCV staining, flow settings for sorting side population cells, and quantitation of radiolabeled DHT retention.