Abstract
Molecular analyses in mammalian meiotic cells have been hindered by the difficulty in isolating stage-specific cell populations, and this is especially true for early meiotic prophase stages (leptotene and zygotene). Here, we describe a method for obtaining cells in different spermatogenic stages from rodents including lepto-zygotene meiocytes at very high purity levels. The procedure includes an approach for the mechanical disaggregation of the testicular tissue, staining with a vital, noncytotoxic dye that is excitable with a blue laser, isolation of the cell populations by flow sorting, and different alternative protocols for the collection of the sorted cells.
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Acknowledgments
This work was supported by Agencia Nacional de Investigación e Innovación (ANII, Uruguay) (Project N° FCE-1-2011-1-6742 to A.G.). The authors thank Federico Santiñaque for his invaluable expertise in the FCM analysis and sorting procedures and for critical reading of the manuscript.
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Geisinger, A., Rodríguez-Casuriaga, R. (2017). Flow Cytometry for the Isolation and Characterization of Rodent Meiocytes. In: Stuart, D. (eds) Meiosis. Methods in Molecular Biology, vol 1471. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-6340-9_11
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DOI: https://doi.org/10.1007/978-1-4939-6340-9_11
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Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-6338-6
Online ISBN: 978-1-4939-6340-9
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