title: Isolation of Undifferentiated Female Germline Cells from Adult Drosophila Ovaries.
abstract: This unit describes a method for isolating undifferentiated, stem cell-like germline cells from adult Drosophila ovaries. Here, we demonstrate that this population of cells can be effectively purified from hand-dissected ovaries in considerably large quantities. Tumor ovaries with expanded populations of undifferentiated germline cells are first removed from fly abdomens and dissociated into a cell suspension with the aid of protease treatment. The target cells, which express Vasa-green fluorescent protein (GFP) fusion protein under the control of the germline-specific vasa promoter, are specifically selected from the suspension via fluorescence-activated cell sorting (FACS). These protocols can be adapted to isolate other cell types from fly ovaries, such as somatic follicle cells or escort cells, by driving GFP expression in the respective target cells.
authors.affiliation: Temasek Life Sciences Laboratory, National University of Singapore, Singapore.|Department of Biological Sciences, National University of Singapore, Singapore.
authors.forename: Robyn Su May
authors.initials: RSM
authors.lastname: Lim
authors.affiliation: Cancer Science Institute of Singapore, National University of Singapore, Singapore.
authors.forename: Motomi
authors.initials: M
authors.lastname: Osato
authors.affiliation: Temasek Life Sciences Laboratory, National University of Singapore, Singapore.|Department of Biological Sciences, National University of Singapore, Singapore.
authors.forename: Toshie
authors.initials: T
authors.lastname: Kai
mesh_terms: D053687:Adult Stem Cells / Q000166:cytology / Q000378:metabolism; D000818:Animals; D030801:Animals, Genetically Modified / Q000235:genetics / Q000378:metabolism; D002469:Cell Separation / Q000379:methods*; D053487:DEAD-box RNA Helicases / Q000235:genetics / Q000378:metabolism; D029721:Drosophila Proteins / Q000235:genetics / Q000378:metabolism; D004331:Drosophila melanogaster; D005260:Female; D049452:Green Fluorescent Proteins / Q000096:biosynthesis / Q000235:genetics; D010053:Ovary / Q000166:cytology / Q000378:metabolism
chemical_list: D029721:Drosophila Proteins; D049452:Green Fluorescent Proteins; C088269:vas protein, Drosophila; D053487:DEAD-box RNA Helicases
keywords: drosophila germline; fluorescence-activated cell sorting (FACS); germline stem cell; isolation