Cultivation and Live Imaging of Drosophila Imaginal Discs.

The ex vivo cultivation and live imaging of wing discs open exciting new research avenues by overcoming the limitations of end-point analysis of fixed tissues. Here we describe how to prepare an optimized wing disc culture medium (WM1) and how to dissect and arrange wing discs for cultivation and live imaging. This protocol enables the study of dynamic phenomena such as cell division and delamination as well as the use of pharmacological compounds and biosensors. Wing discs cultured and imaged as described here, maintain constant levels of proliferation during the first ten hours of culture.

[1]  Konrad Basler,et al.  A high-throughput template for optimizing Drosophila organ culture with response-surface methods , 2013, Development.

[2]  Frank Jülicher,et al.  Local Increases in Mechanical Tension Shape Compartment Boundaries by Biasing Cell Intercalations , 2014, Current Biology.

[3]  D. Montell,et al.  A protocol for culturing Drosophila melanogaster stage 9 egg chambers for live imaging , 2007, Nature Protocols.

[4]  M. Milner,et al.  The growth and differentiation in vitro of leg and wing imaginal disc cells from Drosophila melanogaster , 1988 .

[5]  R. Ueda,et al.  Cell lines from imaginal discs ofDrosophila melanogaster , 1987, In Vitro Cellular & Developmental Biology.

[6]  E. Hafen,et al.  Towards Long Term Cultivation of Drosophila Wing Imaginal Discs In Vitro , 2014, PloS one.

[7]  Luis M. Escudero,et al.  Live imaging of Drosophila imaginal disc development , 2010, Proceedings of the National Academy of Sciences.

[8]  Frank Jülicher,et al.  Increased Cell Bond Tension Governs Cell Sorting at the Drosophila Anteroposterior Compartment Boundary , 2009, Current Biology.

[9]  J. Robb.  MAINTENANCE OF IMAGINAL DISCS OF DROSOPHILA MELANOGASTER IN CHEMICALLY DEFINED MEDIA , 1969, The Journal of cell biology.

[10]  Lennart Kester,et al.  Differential proliferation rates generate patterns of mechanical tension that orient tissue growth , 2013, The EMBO journal.

[11]  Thomas Lecuit,et al.  A global pattern of mechanical stress polarizes cell divisions and cell shape in the growing Drosophila wing disc , 2013, Development.

[12]  A. Shearn,et al.  In vitro growth of imaginal disks from Drosophila melanogaster. , 1977, Science.

[13]  C. Wyss.  Ecdysterone, insulin and fly extract needed for the proliferation of normal Drosophila cells in defined medium. , 1982, Experimental cell research.