Development of an In Vitro Protocol for Drosophila Melanogaster Midgut Cells
Abstract
The potential therapeutic benefits of adult stem cells have created a need to establish a variety of in vitro models to facilitate their study. In the common fruit fly, Drosophila melanogaster, adult stem cell populations have been identified in the nervous, reproductive, and digestive systems. The Drosophila midgut has emerged as a robust model system to study the influence of different signaling pathways and environmental influences on the biology of intestinal stem cells. The ability to culture these cells will facilitate further exploration of these mechanisms and in our case it will enable us to develop toxicology assays and also further continue our work on aging mechanisms. Thus far, no successful cell cultures of the digestive system have been reported. In this study the midgut of Drosophila, an organ very similar in structure and morphology to that of the stomach, small intestine and colon in humans, was used because of the reported presence of adult intestinal stem cell (ISC) population. In order to establish a culture of this tissue, a procedure for its extraction, dissociation, and maintenance in vitro needed to be developed. Utilizing micro-surgery techniques, successful extraction of the targeted tissue was accomplished. A combination of chemical dissociation using a proteolytic enzyme and mechanical dissociation using a micro-homogenizer proved effective in isolating all cell types within the midgut. Standard Drosophila Schneider’s media supplemented with 10%…
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