Given the power of Drosophila genetics, it may seem surprising to discover that many fly researchers are turning to Drosophila cell culture as an experimental system. However, as we will show in this chapter, there are many benefits to be gained by using cell lines as a complement to studies in a tissue and developmental context in the fly. Moreover, one can argue that Drosophila cell culture, in itself, provides an excellent model system for the study of many fundamental questions in molecular and cellular biology. In this review, we offer a summary of techniques that should be useful to researchers in the Drosophila community working with fly cell lines. These include techniques for growing and maintaining cell lines, transient and stable transfection, RNA interference, imaging, immunostaining, fluorescence-activated cell sorting, and for the isolation of RNA and protein from fly cells.