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Isolation of Murine Spermatogenic Cells using a Violet-Excited Cell-Permeable DNA Binding Dye

Article DOI: 10.3791/61666-v • 08:21 min • January 14th, 2021
January 14th, 2021 •
*1,2, *1,2, 3,4, 1,2, 3,4, 5,6, 1,2
1Division of Reproductive Sciences, Division of Developmental Biology, Perinatal Institute, Cincinnati Children's Hospital Medical Center, 2Department of Pediatrics, University of Cincinnati College of Medicine, 3Division of Germ Cell Biology, National Institute for Basic Biology, National Institutes of Natural Sciences, 4Department of Basic Biology, School of Life Science, Graduate University for Advanced Studies (Sokendai), 5Department of Animal Science and Biotechnology, School of Veterinary Medicine, Azabu University, 6Faculty of Science and Technology, Department of Applied Biological Science, Tokyo University of Science
* These authors contributed equally

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Summary
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Here we present a simple and efficient method to isolate live meiotic and post-meiotic germ cells from adult mouse testes. Using a low-cytotoxicity, violet-excited DNA binding dye and fluorescence-activated cell sorting, one can isolate highly enriched spermatogenic cell populations for many downstream applications.

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Isolation Murine Spermatogenic Cells Violet-excited Cell-permeable DNA Binding Dye Subpopulations Spermatocytes Spermatids Adult Mice Molecular Mechanisms Meiosis Spermatogenesis Low Cytotoxicity DNA Binding Dye Excitation Spectrum FACS Orders Flow Cytometry Gating Gating Strategy Protocol Adaptation Testes Harvesting Tunica Albuginea Removal Seminiferous Tubules Dispersion PBS Washing Dissociation Buffer Incubation
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