Biological System: Disc
What are the conditions, tools, and techniques needed to culture and image a developing wing disc in vivo or in vitro over a longer time period?
In vitro and in vivo development of imaginal wing discs enables a precise analysis of the behavior of cells and cellular components in real time. Cultured wing discs allow for high-resolution measurements and perturbation studies using physical, chemical, and genetic tools. Therefore, the development of a reliable culture method to study cell cycle patterns and cell growth is the aim of this WingX subgroup.
The main focus is on the establishment of an in vitro culture system for imaginal discs. Long-term culture of wing imaginal discs in conditions that support normal development in vitro is extremely challenging and has not been reported so far. Systematic investigation of a culture medium that supports development as well as engineering microfluidic culture chambers is a central task. These microfulidic devices will be compatible with light microscopic recording for time-resolved quantitative analyses of development and growth. Complementary to the in vitro culture approach new wing disc cell cultures will be established from embryonic primary cultures.
Another focus lies on in vivo culture methods. In vivo, dissected imaginal discs have been cultivated successfully in the abdomen of adult female flies that are filled with hemolymph. This WingX subgroup establishes a reliable in vivo culture method that permits to address questions of cell cycle patterns and growth.
To study cell cycle patterns and cell growth imaging techniques need to be developed. Within WingX, imaging techniques for live imaging of discs through the larval cuticle have already been established. These techniques will be extended for imaging of in vitro and in vivo cultivated wings discs. In addition, segmentation algorithms allowing to track single cells as well as in vivo cell marker methods, that allow for studying aspects of wing disc growth without direct manipulation, will be evolved.
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