Drosophila Cells in Culture, Second Edition, includes comprehensive coverage of cell lines, methods for creating cell lines, methods for genome engineering, and the use of cell lines for genome wide rNAi screens. This publication summarizes over thirty years of experience in the handling of in vitro cultured Drosophila cells alongside recent methods and functional screens. Early and experienced researchers studying drosophila in developmental biology, genetics, neuroscience, and across the biological and biomedical sciences will benefit from this expert knowledge.