title: Primary cell cultures from Drosophila gastrula embryos.
abstract: Here we describe a method for preparing and culturing primary cells dissociated from Drosophila gastrula embryos. In brief, a large amount of staged embryos from young and healthy flies are collected, sterilized, and then physically dissociated into a single cell suspension using a glass homogenizer. After being plated on culture plates or chamber slides at an appropriate density in culture medium, these cells can further differentiate into several morphologically-distinct cell types, which can be identified by their specific cell markers. Furthermore, we present conditions for treating these cells with double stranded (ds) RNAs to elicit gene knockdown. Efficient RNAi in Drosophila primary cells is accomplished by simply bathing the cells in dsRNA-containing culture medium. The ability to carry out effective RNAi perturbation, together with other molecular, biochemical, cell imaging analyses, will allow a variety of questions to be answered in Drosophila primary cells, especially those related to differentiated muscle and neuronal cells.
authors.affiliation: Department of Genetics, Harvard Medical School.
authors.forename: Norbert
authors.initials: N
authors.lastname: Perrimon
authors.forename: Jonathan
authors.initials: J
authors.lastname: Zirin
authors.forename: Jianwu
authors.initials: J
authors.lastname: Bai
mesh_terms: D000818:Animals; D018929:Cell Culture Techniques / Q000379:methods*; D003470:Culture Media; D004330:Drosophila / Q000166:cytology* / Q000196:embryology / Q000502:physiology; D034622:RNA Interference; D012330:RNA, Double-Stranded / Q000008:administration & dosage / Q000235:genetics
chemical_list: D003470:Culture Media; D012330:RNA, Double-Stranded