Isolation of Undifferentiated Female Germline Cells from Adult Drosophila Ovaries
| Autor: | Robyn Su May Lim, Motomi Osato, Toshie Kai |
|---|---|
| Rok vydání: | 2012 |
| Předmět: |
Aging
Cell type Tissue Fixation Somatic cell Green Fluorescent Proteins Population Cell Separation Biology Germline Green fluorescent protein Animals Genetically Modified DEAD-box RNA Helicases Follicle Suspensions Drosophila Proteins Animals education education.field_of_study Dissection Ovary fungi Cell Differentiation Cell Biology General Medicine Cell sorting Flow Cytometry Molecular biology Fusion protein Adult Stem Cells Drosophila melanogaster Germ Cells Female Peptide Hydrolases Developmental Biology |
| Zdroj: | Current Protocols in Stem Cell Biology |
| ISSN: | 1938-8969 1941-7322 |
| DOI: | 10.1002/9780470151808.sc02e03s22 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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