Flow Cytometry for the Isolation and Characterization of Rodent Meiocytes
Flow Cytometry for the Isolation and Characterization of Rodent Meiocytes
复制标题
DOI：
10.1007/978-1-4939-6340-9_11
复制
发表时间：
2017-01-01
期刊：
影响因子：
--
通讯作者：
Rodriguez-Casuriaga, Rosana
中科院分区：
文献类型：
--
作者：
Geisinger, Adriana;Rodriguez-Casuriaga, Rosana
哺乳动物减数分裂细胞的分子分析因难以分离特定阶段的细胞群而受到阻碍，对于早期减数分裂前期（细线期和合子期）尤其如此。在这里，我们描述了一种从啮齿动物中获得不同生精阶段细胞的方法，包括纯度非常高的细合子性母细胞。该程序包括睾丸组织的机械解聚方法、用可被蓝色激光激发的重要的非细胞毒性染料染色、通过流式分选分离细胞群以及收集分选细胞的不同替代方案。
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.