A high-throughput template for optimizing Drosophila organ culture with response surface methods. Jeremiah J. Zartman^1,2, Simon Restrepo², Konrad Basler². 1) Department of Chemical and Biomolecular Engineering, University of Notre Dame, Notre Dame, IN; 2) Institute of Molecular Life Sciences, University of Zurich, Zurich, Switzerland.

   The Drosophila wing imaginal disc is a key model organ for molecular developmental genetics. Wing disc studies are generally restricted to endpoint analyses of fixed tissues. Recently several studies have relied on limited data from discs cultured in uncharacterized conditions. Systematic efforts toward developing Drosophila organ culture techniques are becoming critical for further progress. Here we designed a multi-tiered, high-throughput pipeline employing design-of-experiment methods to design a culture medium for wing discs. The resulting formula sustains high levels of proliferation for more than twelve hours. This approach results in a statistical model of proliferation as a function of extrinsic growth supplements and identifies synergies that improve insulin-stimulated growth. A more dynamic view of organogenesis emerges from the optimized culture system that highlights important facets of growth: spatio-temporal clustering of cell divisions and cell junction rearrangements. The same approach may prove useful in improving culture conditions for other organ systems.