Isolation of Mammalian Oogonial Stem Cells by Antibody-Based Fluorescence-Activated Cell Sorting
| Autor: | Dori C. Woods, Deanna M. Navaroli, Jonathan L. Tilly |
|---|---|
| Rok vydání: | 2016 |
| Předmět: |
0301 basic medicine
Cell Separation Biology Article Germline Epitope Immunophenotyping Oogonial Stem Cells Flow cytometry DEAD-box RNA Helicases Mice 03 medical and health sciences medicine Animals Humans medicine.diagnostic_test Stem Cells Ovary Cell sorting Flow Cytometry Cell biology Germ Cells 030104 developmental biology medicine.anatomical_structure biology.protein Female Antibody Stem cell Biomarkers Germ cell |
| Zdroj: | Methods in Molecular Biology ISBN: 9781493937936 Methods Mol Biol |
| DOI: | 10.1007/978-1-4939-3795-0_19 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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