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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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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