ArticleJournal of visualized experiments : JoVE2021
Isolation of Murine Spermatogenic Cells using a Violet-Excited Cell-Permeable DNA Binding Dye.
Article in Journal of visualized experiments : JoVE, 2021. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 11 papers.
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Who cites it
11 citing papers in PubMed.
- DAXX directs dual modes of H3.4-to-H3.3 histone replacement in the male germline.Genes & development · 2026Article
- Paternal folate deficiency reveals meiosis as a metabolic sensing window in the male germline.bioRxiv : the preprint server for biology · 2026Article
- CTCF-mediated 3D chromatin sets up the gene expression program in the male germline.Nature structural & molecular biology · 2025Article
- Chromatin remodeler CHD4 establishes chromatin states required for ovarian reserve formation, maintenance and male germ cell survival.Nucleic acids research · 2025Article
- Chromatin remodeler CHD4 establishes chromatin states required for ovarian reserve formation, maintenance, and germ cell survival.bioRxiv : the preprint server for biology · 2024Article
- Atypical heat shock transcription factor HSF5 is critical for male meiotic prophase under non-stress conditions.Nature communications · 2024Article
- ATF7IP2/MCAF2 directs H3K9 methylation and meiotic gene regulation in the male germline.Genes & development · 2024Article
- PRC1 directs PRC2-H3K27me3 deposition to shield adult spermatogonial stem cells from differentiation.Nucleic acids research · 2024Article
- ATF7IP2/MCAF2 directs H3K9 methylation and meiotic gene regulation in the male germline.bioRxiv : the preprint server for biology · 2023Article
- DOT1L promotes spermatid differentiation by regulating expression of genes required for histone-to-protamine replacement.Development (Cambridge, England) · 2023Article
- Active DNA damage response signaling initiates and maintains meiotic sex chromosome inactivation.Nature communications · 2022Article
Corrections and comments
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Authors and funding
7 authors.
Funding
Abstract
Isolation of meiotic spermatocytes is essential to investigate molecular mechanisms underlying meiosis and spermatogenesis. Although there are established cell isolation protocols using Hoechst 33342 staining in combination with fluorescence-activated cell sorting, it requires cell sorters equipped with an ultraviolet laser. Here we describe a cell isolation protocol using the DyeCycle Violet (DCV) stain, a low cytotoxicity DNA binding dye structurally similar to Hoechst 33342. DCV can be excited by both ultraviolet and violet lasers, which improves the flexibility of equipment choice, including a cell sorter not equipped with an ultraviolet laser. Using this protocol, one can isolate three live-cell subpopulations in meiotic prophase I, including leptotene/zygotene, pachytene, and diplotene spermatocytes, as well as post-meiotic round spermatids. We also describe a protocol to prepare single-cell suspension from mouse testes. Overall, the procedure requires a short time to complete (4-5 hours depending on the number of needed cells), which facilitates many downstream applications.
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