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Analysis of cells in culture has made substantial contributions to biological research. The versatility and scale of  in vitro  manipulation and new applications such as high-throughput gene silencing screens ensure the continued importance of cell-culture studies. In comparison to mammalian systems,  Drosophila  cell culture is underdeveloped, primarily because there is no general genetic method for deriving new cell lines. Here we found expression of the conserved oncogene  Ras V12   (a constitutively activated form of  Ras ) profoundly influences the development of primary cultures derived from embryos. The cultures become confluent in about three weeks and can be passaged with great success. The lines have undergone more than 90 population doublings and therefore constitute continuous cell lines. Most lines are composed of spindle-shaped cells of mesodermal type. We tested the use of the method for deriving  Drosophila  cell lines of a specific genotype by establishing cultures from embryos in which the  warts  ( wts ) tumor suppressor gene was targeted. We successfully created several cell lines and found that these differ from controls because they are primarily polyploid. This phenotype likely reflects the known role for the mammalian  wts  counterparts in the tetraploidy checkpoint. We conclude that expression of  Ras V12   is a powerful genetic mechanism to promote proliferation in  Drosophila  primary culture cells and serves as an efficient means to generate continuous cell lines of a given genotype.

Efficient Genetic Method for Establishing Drosophila Cell Lines Unlocks the Potential to Create Lines of Specific Genotypes