title: Isolation of Mammalian Oogonial Stem Cells by Antibody-Based Fluorescence-Activated Cell Sorting.
abstract: The ability to isolate and subsequently culture mitotically active female germ cells from adult ovaries, referred to as either oogonial stem cells (OSCs) or adult female germline stem cells (aFGSCs), has provided a robust system to study female germ cell development under multiple experimental conditions, and in many species. Flow cytometry or fluorescence-activated cell sorting (FACS) is an integral part of many isolation and characterization protocols. Here, we provide methodological details for antibody-based flow cytometric isolation of OSCs using antibodies specific for external epitopes of the proteins Ddx4 or Ifitm3, alone or in combination with the use of fluorescent reporter mice. Beginning with sample preparation, we provide point-by-point instructions to guide researchers on how to isolate OSCs using flow cytometry.
authors.affiliation: Laboratory of Aging and Infertility Research, Northeastern University, 360 Huntington Avenue, Boston, MA, 02115, USA.|Department of Biology, Northeastern University, Boston, MA, 02115, USA.
authors.forename: Deanna M
authors.initials: DM
authors.lastname: Navaroli
authors.affiliation: Laboratory of Aging and Infertility Research, Northeastern University, 360 Huntington Avenue, Boston, MA, 02115, USA. j.tilly@neu.edu.|Department of Biology, Northeastern University, Boston, MA, 02115, USA. j.tilly@neu.edu.
authors.forename: Jonathan L
authors.initials: JL
authors.lastname: Tilly
authors.affiliation: Laboratory of Aging and Infertility Research, Northeastern University, 360 Huntington Avenue, Boston, MA, 02115, USA. d.woods@neu.edu.|Department of Biology, Northeastern University, Boston, MA, 02115, USA. d.woods@neu.edu.
authors.forename: Dori C
authors.initials: DC
authors.lastname: Woods
mesh_terms: D000818:Animals; D015415:Biomarkers; D002469:Cell Separation / Q000379:methods; D053487:DEAD-box RNA Helicases / Q000737:chemistry / Q000276:immunology / Q000378:metabolism; D005260:Female; D005434:Flow Cytometry / Q000379:methods; D005854:Germ Cells / Q000166:cytology* / Q000378:metabolism*; D006801:Humans; D016130:Immunophenotyping; D051379:Mice; D010053:Ovary / Q000166:cytology*; D013234:Stem Cells / Q000166:cytology / Q000378:metabolism
chemical_list: D015415:Biomarkers; D053487:DEAD-box RNA Helicases
keywords: Female germline stem cell; Flow cytometry; Oogonial stem cell; Ovary