Unit

UNIT 2E.3 Isolation of Undifferentiated Female Germline Cells from Adult Drosophila Ovaries

  1. Robyn Su May Lim1,2,
  2. Motomi Osato3,
  3. Toshie Kai1,2

Published Online: 1 AUG 2012

DOI: 10.1002/9780470151808.sc02e03s22

Current Protocols in Stem Cell Biology

Current Protocols in Stem Cell Biology

How to Cite

May Lim, R. S., Osato, M. and Kai, T. 2012. Isolation of Undifferentiated Female Germline Cells from Adult Drosophila Ovaries. Current Protocols in Stem Cell Biology. 22:2E.3.1–2E.3.14.

Author Information

  1. 1

    Temasek Life Sciences Laboratory, National University of Singapore, Singapore

  2. 2

    Department of Biological Sciences, National University of Singapore, Singapore

  3. 3

    Cancer Science Institute of Singapore, National University of Singapore, Singapore

Publication History

  1. Published Online: 1 AUG 2012

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. Curr. Protoc. Stem Cell Biol. 22:2E.3.1-2E.3.14. © 2012 by John Wiley & Sons, Inc.

Keywords:

  • Drosophila germline;
  • germline stem cell;
  • isolation;
  • fluorescence-activated cell sorting (FACS)